48 results on '"U87 glioma cells"'
Search Results
2. ERN1 dependent impact of glutamine and glucose deprivations on the pyruvate dehydrogenase genes expression in glioma cells
- Author
-
Shatokhina Hanna O., Khita Olena O., Minchenko Dmytro O., Tsymbal Dariia O., Luzina Olha R., Danilovskyi Serhiy V., Sliusar Myroslava Y., Levadna Liudmyla O., and Minchenko Oleksandr H.
- Subjects
u87 glioma cells ,ern1 knockdown ,glutamine and glucose deprivations ,pyruvate dehydrogenase ,gene expression ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
Objective. The aim of the present study was to investigate the expression of pyruvate dehydrogenase genes such as PDHA1, PDHB, DLAT, DLD, and PDHX in U87 glioma cells in response to glutamine and glucose deprivations in control glioma cells and endoplasmic reticulum to nucleus signaling 1 (ERN1) knockdown cells, the major endoplasmic reticulum (ER) stress signaling pathway, to find out whether there exists a possible dependence of these important regulatory genes expression on both glutamine and glucose supply as well as ERN1 signaling.
- Published
- 2022
- Full Text
- View/download PDF
3. The impact of glutamine deprivation on the expression of MEIS3, SPAG4, LHX1, LHX2, and LHX6 genes in ERN1 knockdown U87 glioma cells
- Author
-
Krasnytska Dariia A., Khita Olena O., Tsymbal Dariia O., Luzina Olha Y., Cherednychenko Anastasiia A., Kozynkevich Halyna E., Bezrodny Borys H., and Minchenko Dmytro O.
- Subjects
u87 glioma cells ,ern1 knockdown ,glutamine deprivation ,homeobox genes ,mrna expression ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
Objective. The aim of the current study was to investigate the expression of genes encoded homeobox proteins such as MEIS3 (Meis homeobox 3), SPAG4 (sperm associated antigen 4), LHX1 (LIM homeobox 1), LHX2, and LHX6 in U87 glioma cells in response to glutamine deprivation in control glioma cells and cells with knockdown of ERN1 (endoplasmic reticulum to nucleus signaling 1), the major pathway of the endoplasmic reticulum stress signaling, for evaluation of a possible dependence on the expression of these important regulatory genes from glutamine supply and ERN1 signaling.
- Published
- 2022
- Full Text
- View/download PDF
4. ERN1 knockdown modifies the impact of glucose and glutamine deprivations on the expression of EDN1 and its receptors in glioma cells
- Author
-
Minchenko Dmytro O., Khita Olena O., Tsymbal Dariia O., Viletska Yuliia M., Sliusar Myroslava Y., Yefimova Yuliia V., Levadna Liudmyla O., Krasnytska Dariia A., and Minchenko Oleksandr H.
- Subjects
ern1 knockdown ,mrna expression ,edn1 ,ednra ,ednrb ,ece1 ,glucose and glutamine deprivations ,u87 glioma cells ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
Objective. The aim of the present investigation was to study the impact of glucose and gluta-mine deprivations on the expression of genes encoding EDN1 (endothelin-1), its cognate receptors (EDNRA and EDNRB), and ECE1 (endothelin converting enzyme 1) in U87 glioma cells in response to knockdown of ERN1 (endoplasmic reticulum to nucleus signaling 1), a major signaling pathway of endoplasmic reticulum stress, for evaluation of their possible implication in the control of glioma growth through ERN1 and nutrient limitations.
- Published
- 2021
- Full Text
- View/download PDF
5. ERN1 dependent regulation of TMED10, MYL9, SPOCK1, CUL4A and CUL4B genes expression at glucose and glutamine deprivations in U87 glioma cells
- Author
-
O. H. Minchenko, O. S. Hnatiuk, D. O. Tsymbal, Y. M. Viletska, S. V. Danilovskyi, O. V. Halkin, I. V. Kryvdiuk, and O. V. Rudnytska
- Subjects
mrna expression ,tmed10 ,myl9 ,spock1 ,cul4a ,cul4b ,ern1 inhibition ,glucose and glutamine deprivation ,u87 glioma cells ,Biochemistry ,QD415-436 ,Medicine ,Biology (General) ,QH301-705.5 - Abstract
It was shown previously that inhibition of ERN1 (endoplasmic reticulum to nucleus signaling 1) pathway, a central mediator of the unfolded protein response, leads to suppression of tumor growth through down-regulation of key pro-proliferative and up-regulation of tumor suppressor factors and modifies the sensitivity of these genes to glucose and glutamine deprivation. However, the executive mechanisms of ERN1 mediated control of glioma cell proliferation are not yet known. The goal of this study was to estimate the effect of glucose and glutamine deprivations on expression of cancer related genes in glioma U87 cells at ERN1 signaling inhibition for evaluation of their possible significance in ERN1 mediated control of glioma cell proliferation. We have studied the effect of glucose and glutamine deprivations on the expression level of cancer related genes encoding TMED10 (transmembrane p24 trafficking protein 10), MYL9 (myosin, light chain 9, regulatory), SPOCK1 (sparc/osteonectin, cwcv and kazal-like domains proteoglycan 1), CUL4A (cullin 4A), and CUL4B in U87 glioma control cells and cells with ERN1 knockdown. It was shown that at glucose deprivation the expression level of MYL9, SPOCK1 and CUL4B genes was significantly up-regulated in control glioma cells. ERN1 knockdown modified the sensitivity to glucose deprivation of all studied genes except TMED10 gene. At glutamine deprivation the expression of MYL9, CUL4A and CUL4B genes was shown to be up-regulated in control glioma cells. The sensitivity of MYL9, TMED10 and CUL4B gene expression to glutamine deprivation in glioma cells with ERN1 knockdown was significantly modified, while CUL4A and SPOCK1 gene expression did not respond to ERN1 inhibition. The present study demonstrates that glucose and glutamine deprivation affected the expression of the most studied genes in a specific manner and that inhibition of ERN1 signaling preferentially modified their expression at glucose and glutamine deprivation.
- Published
- 2020
- Full Text
- View/download PDF
6. Glucose deprivation affects the expression of genes encoding cAMP-activated protein kinase and related proteins in U87 glioma cells in ERN1 dependent manner
- Author
-
Ratushna Oksana O.
- Subjects
ern1 knockdown ,homeobox genes ,mrna expression ,glucose deprivation ,u87 glioma cells ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
Objective. The aim of this investigation was to study the expression of genes encoding cAMP-activated protein kinase catalytic and regulatory A subunits (PRKACA and PRKAR1A) and related proteins such as cAMP-dependent protein kinase inhibitors A and G (PKIA and PKIG), catalytic subunit A of protein phosphatase 3 (PPP3CA), A-kinase anchoring protein 12 (AKAP12), and praja ring finger ubiquitin ligase 2 (PJA2) in U87 glioma cells in response to glucose deprivation in both control U87 glioma cells and cells with ERN1 (endoplasmic reticulum to nucleus signaling 1) knockdown, the major pathway of the endoplasmic reticulum stress signaling, for evaluation of possible significance of glucose deprivation in ERN1 dependent regulation of glioma growth.
- Published
- 2020
- Full Text
- View/download PDF
7. Insulin receptor substrate 1 gene expression is strongly up-regulated by HSPB8 silencing in U87 glioma cells
- Author
-
Hnatiuk Oksana S., Tsymbal Dariia O., Minchenko Dmytro O., Khita Olena O., Viletska Yulia M., Rundytska Olha V., Kozynkevych Halyna E., Maslak Hanna S., and Minchenko Oleksandr H.
- Subjects
silencing hspb8 ,mrna expression ,irs1 ,homer3 ,perp ,clo1 ,myl9 ,per2 ,gadd45a ,u87 glioma cells ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
Objective. The aim of the present investigation was to study the expression of genes encoding IRS1 (insulin receptor substrate 1) and some other functionally active proteins in U87 glioma cells under silencing of polyfunctional chaperone HSPB8 for evaluation of the possible significance of this protein in intergenic interactions.
- Published
- 2020
- Full Text
- View/download PDF
8. ERN1 knockdown modifies the effect of glucose deprivation on homeobox gene expressions in U87 glioma cells
- Author
-
Tsymbal Dariia O., Minchenko Dmytro O., Khita Olena O., Rudnytska Olha V., Viletska Yulia M., Lahanovska Yulia O., He Qiuxia, Liu Kechun, and Minchenko Oleksandr H.
- Subjects
ern1 knockdown ,homeobox genes ,mrna expression ,glucose deprivation ,u87 glioma cells ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
Objective. The aim of the present investigation was to study the expression of genes encoding homeobox proteins ZEB2 (zinc finger E-box binding homeobox 2), TGIF1 (TGFB induced factor homeobox 1), SPAG4 (sperm associated antigen 4), LHX1 (LIM homeobox 1), LHX2, LHX6, NKX3-1 (NK3 homeobox 1), and PRRX1 (paired related homeobox 1) in U87 glioma cells in response to glucose deprivation in control glioma cells and cells with knockdown of ERN1 (endoplasmic reticulum to nucleus signaling 1), the major pathway of the endoplasmic reticulum stress signaling, for evaluation of it possible significance in the control of glioma growth through ERN1 signaling and chemoresistance.
- Published
- 2020
- Full Text
- View/download PDF
9. Expression of IDE and PITRM1 genes in ERN1 knockdown U87 glioma cells: effect of hypoxia and glucose deprivation
- Author
-
Minchenko Dmytro O., Khita Olena O., Tsymbal Dariia O., Danilovskyi Serhij V., Rudnytska Olha V., Halkin Oleh V., Kryvdiuk Iryna V., Smeshkova Maria V., Yakymchuk Mykhailo M., Bezrodnyi Borys H., and Minchenko Oleksandr H.
- Subjects
ern1 knockdown ,ide ,pitrm1 ,mrna expression ,microrna ,hypoxia ,glucose deprivation ,u87 glioma cells ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
Objective. The aim of the present investigation was to study the expression of genes encoding polyfunctional proteins insulinase (insulin degrading enzyme, IDE) and pitrilysin metallopeptidase 1 (PITRM1) in U87 glioma cells in response to inhibition of endoplasmic reticulum stress signaling mediated by ERN1/IRE1 (endoplasmic reticulum to nucleus signaling 1) for evaluation of their possible significance in the control of metabolism through ERN1 signaling as well as hypoxia, glucose and glutamine deprivations.
- Published
- 2020
- Full Text
- View/download PDF
10. Silencing of NAMPT leads to up-regulation of insulin receptor substrate 1 gene expression in U87 glioma cells
- Author
-
Tsymbal Daria O., Minchenko Dmytro O., Luzina Olena Y., Riabovol Olena O., Danilovskyi Serhiy V., and Minchenko Oleksandr H.
- Subjects
silencing nampt ,mrna expression ,irs1 ,igfbp3 ,per2 ,clu ,bnip3 ,tpd52 ,gadd45a ,u87 glioma cells ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
Objective. The aim of the present study was to investigate the effect of adipokine NAMPT (nicotinamide phosphoribosyltransferase) silencing on the expression of genes encoding IRS1 (insulin receptor substrate 1) and some other proliferation related proteins in U87 glioma cells for evaluation of the possible significance of this adipokine in intergenic interactions.
- Published
- 2020
- Full Text
- View/download PDF
11. Effect of glucose deprivation on the expression of genes encoding glucocorticoid receptor and some related factors in ERN1-knockdown U87 glioma cells
- Author
-
Riabovol Olena O., Tsymbal Dariia O., Minchenko Dmytro O., Lebid-Biletska Kateryna M., Sliusar Myroslava Y., Rudnytska Olha V., and Minchenko Oleksandr H.
- Subjects
glucose deprivation ,mrna expression ,nr3c1 ,nr3c2 ,ahr ,nrip1 ,sgk1 ,sgk3 ,arhgap35 ,nnt ,ern1 inhibition ,u87 glioma cells ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
Objective. The aim of the present study was to examine the effect of glucose deprivation on the expression of genes encoded glucocorticoid receptor (NR3C1) and some related proteins (NR3C2, AHR, NRIP1, NNT, ARHGAP35, SGK1, and SGK3) in U87 glioma cells in response to inhibition of endoplasmic reticulum stress signaling mediated by ERN1/IRE1 (endoplasmic reticulum to nucleus signaling 1/inositol requiring enzyme 1) for evaluation of their possible significance in the control of glioma growth through endoplasmic reticulum stress signaling mediated by IRE1 and glucose deprivation.
- Published
- 2019
- Full Text
- View/download PDF
12. Hypoxic regulation of EDN1, EDNRA, EDNRB, and ECE1 gene expressions in ERN1 knockdown U87 glioma cells
- Author
-
Minchenko Dmytro O., Tsymbal Daria O., Riabovol Olena O., Viletska Yuliia M., Lahanovska Yuliia O., Sliusar Myroslava Y., Bezrodnyi Borys H., and Minchenko Oleksandr H.
- Subjects
hypoxia ,mrna expression ,edn1 ,ednra ,ednrb ,ece1 ,microrna ,ern1 knockdown ,u87 glioma cells ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
Objective. The aim of the present investigation was to study the effect of hypoxia on the expression of genes encoding endothelin-1 (EDN1) and its cognate receptors (EDNRA and EDNRB) as well as endothelin converting enzyme 1 (ECE1) in U87 glioma cells in response to inhibition of endoplasmic reticulum stress signaling mediated by ERN1/IRE1 (endoplasmic reticulum to nucleus signaling 1) for evaluation of their possible significance in the control of glioma growth through ERN1 and hypoxia.
- Published
- 2019
- Full Text
- View/download PDF
13. GLUTAMINE DEPRIVATION AFFECTS THE EXPRESSION OF GENES WHICH CONTROL PYRUVATE DEHYDROGENASE ACTIVITY: THE IMPACT OF ERN1 KNOCKDOWN.
- Author
-
Sliusar, M., Shatokhina, H., Cherednychenko, A., and Minchenko, O.
- Subjects
- *
SLEEP deprivation , *GLUTAMINE , *GENE expression , *REGULATOR genes , *PYRUVATES , *REAL-time control - Abstract
The aim of the current investigation was to study the expression of genes encoded pyruvate dehydrogenase subunits (PDHA1, PDHB, PDHX, DLAT, and DLD) in U87 glioma cells in response to glutamine deprivation in U87 glioma cells in relation to knockdown of ERN1 for evaluation of a possible dependence of the expression of these important regulatory genes from glutamine supply and ERN1 signaling. Methods. The expression of PDHA1, PDHB, PDHX, DLAT, and DLD genes was studied by real-time qPCR in control U87 glioma cells (transfected by vector) and cells with knockdown of ERN1 (transfected by dnERN1) after exposure to glutamine deprivation condition. Total RNA was extracted from glioma cells using TRIZOL reagent. An RNA quantity as well as spectral characteristics was measured using NanoDrop One. For reverse transcription of mRNAs we used Thermo Scientific Verso cDNA Synthesis Kit (Germany). The values of mRNA expressions were normalized to the level of ACTB mRNA and represented as percent of control (100 %). Results. It was shown that the expression level of PDH1, PDHB, DLAT, and DLD genes was down-regulated in control glioma cells treated by glutamine deprivation. At the same time, ERN1 knockdown is suppressed the effect of glutamine deprivation on PDHB and DLD gene expressions in glioma cells, but did not change significantly the impact of glutamine deprivation on the expression of PDHA1, DLAT, and PDHX genes. Conclusions. The results of this investigation demonstrated that the expression of PDH1, PDHB, PDHX, DLAT, and DLD genes was significantly affected by exposure of U87 glioma cells under glutamine deprivation condition and that the effect of glutamine deprivation on the expression of most these genes was modified in cells with knockdown of ERN1, a major signaling pathway of the endoplasmic reticulum stress. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF
14. ERN1 modifies the effect of glutamine deprivation on tumor growth related factors expression in U87 glioma cells
- Author
-
O. H. Minchenko, A. P. Kharkova, O. S. Hnatiuk, O. Y. Luzina, I. V. Kryvdiuk, and A. Y. Kuznetsova
- Subjects
ATF6 ,BIRC5 ,EIF2AK3 ,ERN1 inhibition ,glutamine deprivation ,HSPA5 ,mRNA expression ,RAB5C ,U87 glioma cells ,Biochemistry ,QD415-436 ,Medicine ,Biology (General) ,QH301-705.5 - Abstract
The expression of a subset of genes encoding important tumor growth related factors in U87 glioma cells with ERN1 (endoplasmic reticulum to nucleus signaling 1) loss of function as well as upon glutamine deprivation was studied. It was shown that glutamine deprivation down-regulated the expression level of ATF6 (activating transcription factor 6), EIF2AK3/PERK (eukaryotic translation initiation factor 2 alpha kinase 3), GLO1 (glyoxalase I), BIRC5 (baculoviral IAP repeat-containing 5), and RAB5C (RAB5C, a member of RAS oncogene family) mRNAs in control glioma cells. At the same time, the expression level of HSPB8 (heat shock 22kDa protein 8) and HSPA5/GRP78 (heat shock protein family A (Hsp70) member 5) mRNAs was resistant to glutamine withdrawal in these glioma cells. It was also shown that inhibition of ERN1, which controled cell proliferation and tumor growth, modified the effect of glutamine deprivation on the expression levels of most studied genes in U87 glioma cells: up-regulated the expression of ATF6 and HSPA5 genes and enhanced sensitivity of EIF2AK3 and BIRC5 genes to glutamine withdrawal. Furthermore, the expression of all studied genes, except EIF2AK3, was down-regulated in ERN1 knockdown glioma cells in the presence of glutamine. It was demonstrated that glutamine deprivation affected the expression of most studied genes in ERN1 dependent manner and that these changes possibly contributed to the suppression of glioma growth from cells without ERN1 signaling enzyme function.
- Published
- 2018
- Full Text
- View/download PDF
15. GLUTAMINE DEPRIVATION EFFECT ON DEK, TPD52, BRCA1, ADGRE5, LIF, GNPDA1, AND COL6A1 GENE EXPRESSIONS IN IRE1 KNOCKDOWN U87 GLIOMA CELLS
- Author
-
A. P. Kharkova, Y. A. Garmash, O. S. Hnatiuk, O. Y. Luzina, S. V. Danilovskyi, A.Y. Kuznetsova, and O. H. Minchenko
- Subjects
mRNA expression ,DEK ,BRCA1 ,COL6A1 ,ADGRE5 ,GNPDA1 genes ,glutamine deprivation ,IRE1 knockdown ,U87 glioma cells ,Biotechnology ,TP248.13-248.65 - Abstract
To study effect of glutamine deprivation on the expression of genes encoding the key proliferation associated factors on a relation to inhibition of inositol requiring enzyme-1 IRE1 in U87 glioma cells was the aim of the research. It was shown that glutamine withdrawal down-regulated the expression of DEK, BRCA1, LIF, and COL6A1 genes in control glioma cells (transfected by empty vector), up-regulated ADGRE5 gene expression, and did not significantly change the expression of TPD52 and GNPDA. Inhibition of ІRE1 signaling enzyme activity modified the effect of glutamine deprivation on the expression of TPD52, BRCA1, LIF, DEK, ADGRE5, and COL6A1 genes: introduces the effect of glutamine deprivation on TPD52 and GNPDA1, reduced – on COL6A1, and enhanced – on ADGRE5, DEK, and BRCA1 in U87 glioma cells. Therefore, glutamine deprivation affect the expression level of most studied genes in U87 glioma cells in relation to the functional activity of IRE1 signaling enzyme, which is responsible for control of cell proliferation and glioma growth.
- Published
- 2017
- Full Text
- View/download PDF
16. Expression of ubiquitin specific peptidase and ATG7 genes in U87 glioma cells upon glutamine deprivation
- Author
-
O. V. Halkin, D. O. Minchenko, О. O. Riabovol, V. V. Telychko, О. O. Ratushna, and O. H. Minchenko
- Subjects
ATG7 ,glutamine deprivation ,IRE1 inhibition ,mRNA expression ,U87 glioma cells ,USPs ,Biochemistry ,QD415-436 ,Medicine ,Biology (General) ,QH301-705.5 - Abstract
We have studied the effect of glutamine deprivation on the expression of genes encoding for ubiquitin specific peptidases (USP) and ubiquitin activating enzyme E1-like protein/autophagy related 7 (GSA7/ATG7) in U87 glioma cells in relation to inhibition of inositol requiring enzyme-1 (IRE1). It was shown that exposure of control glioma cells (transfected by empty vector) upon glutamine deprivation led to suppression of USP1 and ATG7 mRNA expression and up-regulated USP25 mRNA. At the same time, glutamine deprivation did not significantly change USP4, USP10, USP14, and USP22 gene expressions in these cells. Inhibition of ІRE1 signaling enzyme function in U87 glioma cells increased effect of glutamine deprivation on the expression of USP1 gene and introduced sensitivity of USP4 and USP14 genes to this condition. Therefore, glutamine deprivation affected the expression level of most studied genes in gene specific manner in relation to the functional activity of IRE1 enzyme, a central mediator of endoplasmic reticulum stress, which controls cell proliferation and tumor growth.
- Published
- 2017
- Full Text
- View/download PDF
17. Expression of tumor growth related genes in IRE1 knockdown U87 glioma cells: effect of hypoxia
- Author
-
O. H. Minchenko, O. Y. Luzina, O. S. Hnatiuk, D. O. Minchenko, I. A. Garmash, and O. O. Ratushna
- Subjects
BRCA1 ,COL6A1 ,HOMER3 ,hypoxia ,IRE1 knockdown ,mRNA expression ,TPD52 ,U87 glioma cells ,Biochemistry ,QD415-436 ,Medicine ,Biology (General) ,QH301-705.5 - Abstract
We have studied the effect of IRE1 signaling enzyme knockdown as well as hypoxia on the expression of genes encoding the important tumor growth related proteins (BRCA1, DEK, BCL2L1, COL6A1, TPD52, HOMER3, and GNPDA1) in U87 glioma cells. It was shown that the expression level of breast cancer 1 early onset (BRCA1) and tumor protein D52 (TPD52) mRNAs are strongly up-regulated in U87 glioma cells by down-regulation of IRE1 expression in comparison with the control cells. At the same time the expression level of collagen, type VI, alpha 1 (COL6A1), DEK oncogene (DEK), glucosamine-6-phosphate deaminase 1 (GNPDA1) and homer homolog 3 (HOMER3) was significantly down-regulated in glioma cells under these experimental conditions. It was also shown that hypoxia up-regulated the expression level of COL6A1 and TPD52 mRNAs and down-regulated – BRCA1, DEK, and GNPDA1 mRNAs in control glioma cells and that down-regulation of IRE1, which control cell proliferation and tumor growth, modified the effect of hypoxia on the expression of COL6A1, DEK, BCL2L1, HOMER3, and GNPDA1 genes. The present study demonstrated that hypoxia affected the expression of most studied genes in IRE1-dependent manner.
- Published
- 2017
- Full Text
- View/download PDF
18. Hypoxic regulation of the expression of genes encoded estrogen related proteins in U87 glioma cells: eff ect of IRE1 inhibition
- Author
-
Do Minchenko, Oo Riabovol, Oo Ratushna, and Oh Minchenko
- Subjects
ire1 inhibition ,hypoxia ,mrna expression ,nrip1 ,ebbp ,esrra ,e2ig5 ,pgrmc2 ,slc39a6 ,u87 glioma cells ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
Objective. The aim of the present study was to examine the effect of inhibition of endoplasmic reticulum stress signaling, mediated by IRE1 (inositol requiring enzyme 1), which is a central mediator of the unfolded protein response on the expression of genes encoded estrogen related proteins (NRIP1/RIP140, TRIM16/EBBP, ESRRA/NR3B1, FAM162A/E2IG5, PGRMC2/PMBP, and SLC39A6/LIV-1) and their hypoxic regulation in U87 glioma cells for evaluation of their possible significance in the control of glioma cells proliferation.
- Published
- 2017
- Full Text
- View/download PDF
19. EXPRESSION OF UBIQUITIN SPECIFIC PEPTIDASE GENES IN IRE1 KNOCKDOWN U87 GLIOMA CELLS UPON GLUCOSE DEPRIVATION
- Author
-
O. H. Minchenko, O. O. Riabovol, O. V. Halkin, S. V. Danilovskyi, D. O. Minchenko, and O. O. Ratushna
- Subjects
mRNA expression ,USP genes ,IRE1 inhibition ,glucose deprivation ,U87 glioma cells ,Biotechnology ,TP248.13-248.65 - Abstract
We have studied the effect of glucose deprivation on the expression of genes encoding for ubiquitin specific peptidases — USP and autophagy related 7 ATG7 in U87 glioma cells in relation to inhibition of inositol requiring enzyme-1 (IRE1). It was shown that glucose deprivation was downregulated the expression of USP1 and USP10 genes and up-regulated USP4 and USP25 genes in control (transfected by empty vector) glioma cells. At the same time, the expression level of USP14, USP22, and ATG7 genes in these cells did not significantly change upon glucose deprivation condition. Inhibition of ІRE1 signaling enzyme function in U87 glioma cells modified effect of glucose deprivation on the expression of most studied genes. Therefore, glucose deprivation affected the expression level of most ubiquitin specific peptidases genes in relation to the functional activity of IRE1 enzyme, which controls cell proliferation and tumor growth as a central mediator of endoplasmic reticulum stress.
- Published
- 2016
- Full Text
- View/download PDF
20. Inhibition of IRE1 signaling affects the expression of genes encoded glucocorticoid receptor and some related factors and their hypoxic regulation in U87 glioma cells
- Author
-
Minchenko D.O., Riabovol O.O., Tsymbal D.O., Ratushna O.O., and Minchenko O.H.
- Subjects
ire1 inhibition ,mrna expression ,nr3c1 ,sgk1 ,sgk3 ,ncoa2 ,arhgap35 ,nnt ,hypoxia ,u87 glioma cells ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
Objective. The aim of the present investigation was to examine the effect of inhibition of endoplasmic reticulum stress signaling, mediated by IRE1 (inositol requiring enzyme 1), which is a central mediator of the unfolded protein response on the expression of genes encoding glucocorticoid receptor (NR3C1) and some related proteins (SGK1, SGK3, NCOA1, NCOA2, ARHGAP35, NNT) and their hypoxic regulation in U87 glioma cells for evaluation of their possible significance in the control of the glioma growth.
- Published
- 2016
- Full Text
- View/download PDF
21. Effect of hypoxia on the expression of genes encoding insulin-like growth factors and some related proteins in U87 glioma cells without IRE1 function
- Author
-
Minchenko Dmytro O., Kharkova A. P., Halkin O. V., Karbovskyi L. L., and Minchenko O. H.
- Subjects
hypoxia ,igf1 ,igf1r ,igfbp4 ,stc2 ,mrna expression ,ire1 inhibition ,u87 glioma cells ,Diseases of the endocrine glands. Clinical endocrinology ,RC648-665 - Abstract
Objective. The aim of the present study was to investigate the effect of hypoxia on the expression of genes encoding insulin-like growth factors (IGF1 and IGF2), their receptor (IGF1R), binding protein-4 (IGFBP4), and stanniocalcin 2 (STC2) in U87 glioma cells in relation to inhibition of endoplasmic reticulum stress signaling mediated by IRE1 (inositol requiring enzyme 1) for evaluation of their possible significance in the control of tumor growth.
- Published
- 2016
- Full Text
- View/download PDF
22. GLUTAMINE DEPRIVATION EFFECT ON DEK, TPD52, BRCA1, ADGRE5, LIF, GNPDA1, AND COL6A1 GENE EXPRESSIONS IN IRE1 KNOCKDOWN U87 GLIOMA CELLS.
- Author
-
Kharkova, A. P., Garmash, Y. A., Hnatiuk, O. S., Luzina, O. Y., Danilovskyi, S. V., Kuznetsova, A.Y., and Minchenko, O. H.
- Subjects
- *
GLUTAMINE , *GENE expression , *INOSITOL , *GLIOMAS , *CELL proliferation - Abstract
To study effect of glutamine deprivation on the expression of genes encoding the key proliferation associated factors on a relation to inhibition of inositol requiring enzyme-1 IRE1 in U87 glioma cells was the aim of the research. It was shown that glutamine deprivation down-regulated the expression of DEK, BRCA1, LIF, and COL6A1 genes in control glioma cells (transfected by empty vector), up-regulated ADGRE5 gene expression, and did not significantly change the expression of TPD52 and GNPDA1. Inhibition of ІRE1 signaling enzyme activity modified the effect of glutamine deprivation on the expression of TPD52, BRCA1, LIF, DEK, ADGRE5, and COL6A1 genes: introduces the effect of glutamine deprivation on TPD52 and GNPDA1, reduced — on COL6A1, and enhanced — on ADGRE5, DEK, and BRCA1 in U87 glioma cells. Therefore, glutamine deprivation affect the expression level of most studied genes in U87 glioma cells in relation to the functional activity of IRE1 signaling enzyme, which is responsible for control of cell proliferation and glioma growth. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
23. ERN1 knockdown modifies the impact of glucose and glutamine deprivations on the expression of EDN1 and its receptors in glioma cells
- Author
-
Yuliia V. Yefimova, Liudmyla O Levadna, Olena O. Khita, Yuliia M. Viletska, Dmytro O. Minchenko, Dariia O. Tsymbal, Dariia A Krasnytska, Oleksandr H. Minchenko, and Myroslava Y. Sliusar
- Subjects
0301 basic medicine ,Endothelin converting enzyme 1 ,Endocrinology, Diabetes and Metabolism ,Glutamine ,Gene Expression ,Endothelin-Converting Enzymes ,Protein Serine-Threonine Kinases ,Diseases of the endocrine glands. Clinical endocrinology ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,ece1 ,Glioma ,Cell Line, Tumor ,Gene expression ,Endoribonucleases ,medicine ,Humans ,ednra ,RNA, Messenger ,ednrb ,education ,Receptor ,Gene knockdown ,education.field_of_study ,Endothelin-1 ,Chemistry ,u87 glioma cells ,ern1 knockdown ,edn1 ,medicine.disease ,Receptor, Endothelin A ,RC648-665 ,Receptor, Endothelin B ,Cell biology ,ERN1 ,030104 developmental biology ,Glucose ,030220 oncology & carcinogenesis ,Gene Knockdown Techniques ,mrna expression ,Signal transduction ,glucose and glutamine deprivations - Abstract
Objective. The aim of the present investigation was to study the impact of glucose and gluta-mine deprivations on the expression of genes encoding EDN1 (endothelin-1), its cognate receptors (EDNRA and EDNRB), and ECE1 (endothelin converting enzyme 1) in U87 glioma cells in response to knockdown of ERN1 (endoplasmic reticulum to nucleus signaling 1), a major signaling pathway of endoplasmic reticulum stress, for evaluation of their possible implication in the control of glioma growth through ERN1 and nutrient limitations. Methods. The expression level of EDN1, its receptors and converting enzyme 1 in control U87 glioma cells and cells with knockdown of ERN1 treated by glucose or glutamine deprivation by quantitative polymerase chain reaction was studied. Results. We showed that the expression level of EDN1 and ECE1 genes was significantly up-regulated in control U87 glioma cells exposure under glucose deprivation condition in comparison with the glioma cells, growing in regular glucose containing medium. We also observed up-regulation of ECE1 gene expression in U87 glioma cells exposure under glutamine deprivation as well as down-regulation of the expression of EDN1 and EDNRA mRNA, being more significant for EDN1. Furthermore, the knockdown of ERN1 signaling enzyme function significantly modified the response of most studied gene expressions to glucose and glutamine deprivation conditions. Thus, the ERN1 knockdown led to a strong suppression of EDN1 gene expression under glucose deprivation, but did not change the effect of glutamine deprivation on its expression. At the same time, the knockdown of ERN1 signaling introduced the sensitivity of EDNRB gene to both glucose and glutamine deprivations as well as completely removed the impact of glucose deprivation on the expression of ECE1 gene. Conclusions. The results of this study demonstrated that the expression of endothelin-1, its receptors, and ECE1 genes is preferentially sensitive to glucose and glutamine deprivations in gene specific manner and that knockdown of ERN1 significantly modified the expression of EDN1, EDNRB, and ECE1 genes in U87 glioma cells. It is possible that the observed changes in the expression of studied genes under nutrient deprivation may contribute to the suppressive effect of ERN1 knockdown on glioma cell proliferation and invasiveness.
- Published
- 2021
24. Insulin receptor substrate 1 gene expression is strongly up-regulated by HSPB8 silencing in U87 glioma cells
- Author
-
Oksana S. Hnatiuk, Dariia O. Tsymbal, Dmytro O. Minchenko, Olha V. Rundytska, Yulia M. Viletska, Olena O. Khita, Halyna E. Kozynkevych, H. S. Maslak, and Oleksandr H. Minchenko
- Subjects
0301 basic medicine ,Endocrinology, Diabetes and Metabolism ,myl9 ,medicine.disease_cause ,Diseases of the endocrine glands. Clinical endocrinology ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,irs1 ,Downregulation and upregulation ,clo1 ,gadd45a ,Cell Line, Tumor ,Glioma ,medicine ,Humans ,Gene silencing ,Gene Silencing ,RNA, Messenger ,neoplasms ,Gene ,Heat-Shock Proteins ,Messenger RNA ,u87 glioma cells ,RC648-665 ,medicine.disease ,nervous system diseases ,Up-Regulation ,IRS1 ,Cell biology ,silencing hspb8 ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,030220 oncology & carcinogenesis ,homer3 ,Insulin Receptor Substrate Proteins ,per2 ,mrna expression ,perp ,Carcinogenesis ,GADD45A ,Molecular Chaperones - Abstract
Objective. The aim of the present investigation was to study the expression of genes encoding IRS1 (insulin receptor substrate 1) and some other functionally active proteins in U87 glioma cells under silencing of polyfunctional chaperone HSPB8 for evaluation of the possible significance of this protein in intergenic interactions. Methods. Silencing of HSPB8 mRNA was introduced by HSPB8 specific siRNA. The expression level of HSPB8, IRS1, HK2, GLO1, HOMER3, MYL9, NAMPT, PER2, PERP, GADD45A, and DEK genes was studied in U87 glioma cells by quantitative polymerase chain reaction. Results. It was shown that silencing of HSPB8 mRNA by specific to HSPB8 siRNA led to a strong down-regulation of this mRNA and significant modification of the expression of IRS1 and many other genes in glioma cells: strong up-regulated of HOMER3, GLO1, and PERP and down-regulated of MYL9, NAMPT, PER2, GADD45A, and DEK gene expressions. At the same time, no significant changes were detected in the expression of HK2 gene in glioma cells treated by siRNA, specific to HSPB8. Moreover, the silencing of HSPB8 mRNA enhanced the glioma cells proliferation rate. Conclusions. Results of this investigation demonstrated that silencing of HSPB8 mRNA affected the expression of IRS1 gene as well as many other genes encoding tumor growth related proteins. It is possible that the dysregulation of most of the studied genes in glioma cells after silencing of HSPB8 is reflected by a complex of intergenic interactions and that this polyfunctional chaperone is an important factor for the stability of genome function and regulatory mechanisms contributing to the tumorigenesis control.
- Published
- 2020
25. ERN1 knockdown modifies the effect of glucose deprivation on homeobox gene expressions in U87 glioma cells
- Author
-
Dariia O. Tsymbal, Yulia O. Lahanovska, Dmytro O. Minchenko, Olha V. Rudnytska, Oleksandr H. Minchenko, Kechun Liu, Yulia M. Viletska, Qiuxia He, and Olena O. Khita
- Subjects
0301 basic medicine ,Endocrinology, Diabetes and Metabolism ,Protein Serine-Threonine Kinases ,Diseases of the endocrine glands. Clinical endocrinology ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Glioma ,Cell Line, Tumor ,Endoribonucleases ,medicine ,Humans ,Gene ,Homeodomain Proteins ,Gene knockdown ,Brain Neoplasms ,u87 glioma cells ,Endoplasmic reticulum ,glucose deprivation ,Genes, Homeobox ,ern1 knockdown ,Transfection ,homeobox genes ,medicine.disease ,RC648-665 ,Cell Hypoxia ,Cell biology ,ERN1 ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,Real-time polymerase chain reaction ,Glucose ,030220 oncology & carcinogenesis ,Gene Knockdown Techniques ,embryonic structures ,Homeobox ,mrna expression ,Energy Metabolism ,Signal Transduction - Abstract
Objective. The aim of the present investigation was to study the expression of genes encoding homeobox proteins ZEB2 (zinc finger E-box binding homeobox 2), TGIF1 (TGFB induced factor homeobox 1), SPAG4 (sperm associated antigen 4), LHX1 (LIM homeobox 1), LHX2, LHX6, NKX3-1 (NK3 homeobox 1), and PRRX1 (paired related homeobox 1) in U87 glioma cells in response to glucose deprivation in control glioma cells and cells with knockdown of ERN1 (endoplasmic reticulum to nucleus signaling 1), the major pathway of the endoplasmic reticulum stress signaling, for evaluation of it possible significance in the control of glioma growth through ERN1 signaling and chemoresistance. Methods. The expression level of homeobox family genes was studied in control (transfected by vector) and ERN1 knockdown U87 glioma cells under glucose deprivation condition by real-time quantitative polymerase chain reaction. Results. It was shown that the expression level of ZEB2, TGIF1, PRRX1, and LHX6 genes was up-regulated in control glioma cells treated by glucose deprivation. At the same time, the expression level of three other genes (NKX3-1, LHX1, and LHX2) was down-regulated. Furthermore, ERN1 knockdown of glioma cells significantly modified the effect glucose deprivation condition on the expression almost all studied genes. Thus, treatment of glioma cells without ERN1 enzymatic activity by glucose deprivation condition lead to down-regulation of the expression level of ZEB2 and SPAG4 as well as to more significant up-regulation of PRRX1 and TGIF1 genes. Moreover, the expression of LHX6 and NKX3-1 genes lost their sensitivity to glucose deprivation but LHX1 and LHX2 genes did not change it significantly. Conclusions. The results of this investigation demonstrate that ERN1 knockdown significantly modifies the sensitivity of most studied homeobox gene expressions to glucose deprivation condition and that these changes are a result of complex interaction of variable endoplasmic reticulum stress related and unrelated regulatory factors and contributed to glioma cell growth and possibly to their chemoresistance.
- Published
- 2020
26. Expression of IDE and PITRM1 genes in ERN1 knockdown U87 glioma cells: effect of hypoxia and glucose deprivation
- Author
-
Olena O. Khita, Olha V. Rudnytska, Oleksandr H. Minchenko, Oleh V. Halkin, Borys H. Bezrodnyi, Mykhailo M. Yakymchuk, Serhij V. Danilovskyi, Iryna V. Kryvdiuk, Dariia O. Tsymbal, Maria V. Smeshkova, and Dmytro O. Minchenko
- Subjects
0301 basic medicine ,microrna ,Endocrinology, Diabetes and Metabolism ,Endoribonuclease activity ,Protein Serine-Threonine Kinases ,Insulysin ,Diseases of the endocrine glands. Clinical endocrinology ,pitrm1 ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Cell Line, Tumor ,Endoribonucleases ,microRNA ,Insulin-degrading enzyme ,Humans ,Protein kinase A ,Messenger RNA ,Gene knockdown ,Brain Neoplasms ,ide ,hypoxia ,u87 glioma cells ,Endoplasmic reticulum ,glucose deprivation ,Metalloendopeptidases ,ern1 knockdown ,Glioma ,Endoplasmic Reticulum Stress ,RC648-665 ,Cell Hypoxia ,Cell biology ,Gene Expression Regulation, Neoplastic ,ERN1 ,Glucose ,030104 developmental biology ,Gene Knockdown Techniques ,mrna expression ,030217 neurology & neurosurgery ,Signal Transduction - Abstract
Objective. The aim of the present investigation was to study the expression of genes encoding polyfunctional proteins insulinase (insulin degrading enzyme, IDE) and pitrilysin metallopeptidase 1 (PITRM1) in U87 glioma cells in response to inhibition of endoplasmic reticulum stress signaling mediated by ERN1/IRE1 (endoplasmic reticulum to nucleus signaling 1) for evaluation of their possible significance in the control of metabolism through ERN1 signaling as well as hypoxia, glucose and glutamine deprivations. Methods. The expression level of IDE and PITRM1 genes was studied in control and ERN1 knockdown U87 glioma cells under glucose and glutamine deprivations as well as hypoxia by quantitative polymerase chain reaction. Results. It was found that the expression level of IDE and PITRM1 genes was down-regulated in ERN1 knockdown (without ERN1 protein kinase and endoribonuclease activity) glioma cells in comparison with the control glioma cells, being more significant for PITRM1 gene. We also found up-regulation of microRNA MIR7-2 and MIRLET7A2, which have specific binding sites in 3’-untranslated region of IDE and PITRM1 mRNAs, correspondingly, and can participate in posttranscriptional regulation of these mRNA expressions. Only inhibition of ERN1 endoribonuclease did not change significantly the expression of IDE and PITRM1 genes in glioma cells. The expression of IDE and PITRM1 genes is preferentially regulated by ERN1 protein kinase. We also showed that hypoxia down-regulated the expression of IDE and PITRM1 genes and that knockdown of ERN1 signaling enzyme function modified the response of these gene expressions to hypoxia. Glucose deprivation increased the expression level of IDE and PITRM1 genes, but ERN1 knockdown enhanced only the effect of glucose deprivation on PITRM1 gene expression. Glutamine deprivation did not affect the expression of IDE gene in both types of glioma cells, but up-regulated PITRM1 gene and this up-regulation was stronger in ERN1 knockdown cells. Conclusions. Results of this investigation demonstrate that ERN1 knockdown significantly decreases the expression of IDE and PITRM1 genes by ERN1 protein kinase mediated mechanism. The expression of both studied genes was sensitive to hypoxia as well as glucose deprivation and dependent on ERN1 signaling in gene-specific manner. It is possible that the level of these genes expression under hypoxia and glucose deprivation is a result of complex interaction of variable endoplasmic reticulum stress related and unrelated regulatory factors and contributed to the control of the cell metabolism.
- Published
- 2020
27. Silencing of NAMPT leads to up-regulation of insulin receptor substrate 1 gene expression in U87 glioma cells
- Author
-
Dmytro O. Minchenko, O O Riabovol, Olena Y. Luzina, Serhiy V. Danilovskyi, Daria O. Tsymbal, and Oleksandr H. Minchenko
- Subjects
0301 basic medicine ,Endocrinology, Diabetes and Metabolism ,Nicotinamide phosphoribosyltransferase ,Gene Expression ,Cell Cycle Proteins ,chemistry.chemical_compound ,0302 clinical medicine ,Endocrinology ,gadd45a ,RNA, Small Interfering ,Nicotinamide Phosphoribosyltransferase ,clu ,Glioma ,Period Circadian Proteins ,Neoplasm Proteins ,Up-Regulation ,Gene Expression Regulation, Neoplastic ,bnip3 ,030220 oncology & carcinogenesis ,Cytokines ,per2 ,mrna expression ,igfbp3 ,GADD45A ,silencing nampt ,MKI67 Gene ,Down-Regulation ,Diseases of the endocrine glands. Clinical endocrinology ,03 medical and health sciences ,Downregulation and upregulation ,irs1 ,Cell Line, Tumor ,Proto-Oncogene Proteins ,medicine ,Gene silencing ,Humans ,tpd52 ,RNA, Messenger ,neoplasms ,Cell Proliferation ,Messenger RNA ,u87 glioma cells ,Membrane Proteins ,medicine.disease ,RC648-665 ,IRS1 ,nervous system diseases ,030104 developmental biology ,Clusterin ,Insulin-Like Growth Factor Binding Protein 3 ,chemistry ,Cancer research ,Insulin Receptor Substrate Proteins - Abstract
Objective. The aim of the present study was to investigate the effect of adipokine NAMPT (nicotinamide phosphoribosyltransferase) silencing on the expression of genes encoding IRS1 (insulin receptor substrate 1) and some other proliferation related proteins in U87 glioma cells for evaluation of the possible significance of this adipokine in intergenic interactions. Methods. The silencing of NAMPT mRNA was introduced by NAMPT specific siRNA. The expression level of NAMPT, IGFBP3, IRS1, HK2, PER2, CLU, BNIP3, TPD52, GADD45A, and MKI67 genes was studied in U87 glioma cells by quantitative polymerase chain reaction. Anti-visfatin antibody was used for detection of NAMPT protein by Western-blot analysis. Results. It was shown that the silencing of NAMPT mRNA led to a strong down-regulation of NAMPT protein and significant modification of the expression of IRS1, IGFBP3, CLU, HK2, BNIP3, and MKI67 genes in glioma cells and a strong up-regulation of IGFBP3 and IRS1 and down-regulation of CLU, BNIP3, HK2, and MKI67 gene expressions. At the same time, no significant changes were detected in the expression of GADD45A, PER2, and TPD52 genes in glioma cells treated by siRNA specific to NAMPT. Furthermore, the silencing of NAMPT mRNA suppressed the glioma cell proliferation. Conclusions. Results of this investigation demonstrated that silencing of NAMPT mRNA with corresponding down-regulation of NAMPT protein and suppression of the glioma cell proliferation affected the expression of IRS1 gene as well as many other genes encoding the proliferation related proteins. It is possible that dysregulation of most of the studied genes in glioma cells after silencing of NAMPT is reflected by a complex of intergenic interactions and that NAMPT is an important factor for genome stability and regulatory mechanisms contributing to the control of glioma cell metabolism and proliferation.
- Published
- 2020
28. IRE1 KNOCKDOWN MODIFIES THE GLUTAMINE AND GLUCOSE DEPRIVATION EFFECT ON THE EXPRESSION OF NUCLEAR GENES ENCODING MITOCHONDRIAL PROTEINS IN U87 GLIOMA CELLS.
- Author
-
Riabovol, O. O., Tsymbal, D. O., Minchenko, D. O., Ratushna, O. O., and Minchenko, O. H.
- Subjects
- *
PROTEIN kinases , *PHYSIOLOGICAL effects of glutamine , *PHYSIOLOGICAL effects of glucose , *MITOCHONDRIAL proteins , *GLIOMAS - Abstract
We have studied the glucose and glutamine deprivation effect on the expression of nuclear genes encoding mitochondrial proteins in U87 glioma cells in relation to inhibition of inositol requiring enzyme-1 (IRE1). It was shown that glutamine deprivation down-regulated the expression of mitochondrial (NADP+)-dependent isocitrate dehydrogenase 2 (IDH2), malic enzyme 2 (ME2), mitochondrial aspartate aminotransferase (GOT2), and subunit B of succinate dehydrogenase (SDHB) genes in control glioma cells in gene specific manner. At the same time, the expression level of malate dehydrogenase 2 (MDH2) and subunit D of succinate dehydrogenase (SDHD) genes in these cells was not changed upon glutamine deprivation. It was also shown that inhibition of ІRE1 signaling enzyme function in U87 glioma cells modified the glutamine deprivation effect on the expression of all studied genes. Furthermore, the expression of the majority of studied genes was resistant to glucose deprivation, except IDH2 and SDHB genes, which expression levels were slightly down-regulated. Inhibition of IRE1 modified the effect of glucose deprivation on ME2, SDHB, SDHD, and GOT2 genes expression. Therefore, glucose and glutamine deprivation affected the expression level of the majority of nuclear genes encoding mitochondrial proteins in relation to the functional activity of IRE1 enzyme, which is a central mediator of endoplasmic reticulum stress and controls cell proliferation and tumor growth. [ABSTRACT FROM AUTHOR]
- Published
- 2016
- Full Text
- View/download PDF
29. Effect of glucose deprivation on the expression of genes encoding glucocorticoid receptor and some related factors in ERN1-knockdown U87 glioma cells
- Author
-
Oleksandr H. Minchenko, Myroslava Y. Sliusar, Olha V. Rudnytska, Dariia O. Tsymbal, O O Riabovol, Kateryna M. Lebid-Biletska, and Dmytro O. Minchenko
- Subjects
0301 basic medicine ,nr3c1 ,Endocrinology, Diabetes and Metabolism ,nr3c2 ,0302 clinical medicine ,Endocrinology ,Glucocorticoid receptor ,Gene expression ,Guanine Nucleotide Exchange Factors ,Gene knockdown ,Brain Neoplasms ,glucose deprivation ,ahr ,Transfection ,Glioma ,nnt ,Cell biology ,Nuclear Receptor Interacting Protein 1 ,Gene Expression Regulation, Neoplastic ,Real-time polymerase chain reaction ,030220 oncology & carcinogenesis ,Gene Knockdown Techniques ,mrna expression ,Signal Transduction ,sgk3 ,sgk1 ,Protein Serine-Threonine Kinases ,nrip1 ,NADP Transhydrogenase, AB-Specific ,Diseases of the endocrine glands. Clinical endocrinology ,Immediate-Early Proteins ,Mitochondrial Proteins ,arhgap35 ,03 medical and health sciences ,Receptors, Glucocorticoid ,Cell Line, Tumor ,Endoribonucleases ,medicine ,Humans ,ern1 inhibition ,Cell Proliferation ,u87 glioma cells ,Endoplasmic reticulum ,Aryl Hydrocarbon Receptor Nuclear Translocator ,medicine.disease ,RC648-665 ,ERN1 ,Repressor Proteins ,030104 developmental biology ,Glucose - Abstract
Objective. The aim of the present study was to examine the effect of glucose deprivation on the expression of genes encoded glucocorticoid receptor (NR3C1) and some related proteins (NR3C2, AHR, NRIP1, NNT, ARHGAP35, SGK1, and SGK3) in U87 glioma cells in response to inhibition of endoplasmic reticulum stress signaling mediated by ERN1/IRE1 (endoplasmic reticulum to nucleus signaling 1/inositol requiring enzyme 1) for evaluation of their possible significance in the control of glioma growth through endoplasmic reticulum stress signaling mediated by IRE1 and glucose deprivation. Methods. The expression of NR3C1, NR3C2, AHR, NRIP1, NNT, ARHGAP35, SGK1, and SGK3 genes in U87 glioma cells transfected by empty vector pcDNA3.1 (control cells) and cells without ERN1 signaling enzyme function (transfected by dnERN1) under glucose deprivation was studied by real time quantitative polymerase chain reaction. Results. It was shown that the expression level of NR3C2, AHR, SGK1, SGK3, and NNT genes was up-regulated in control U87 glioma cells under glucose deprivation condition in comparison with the control cells growing with glucose. At the same time, the expression of NRIP1 gene is down-regulated in these glioma cells under glucose deprivation, but NR3C1 and ARHGAP35 genes was resistant to this experimental condition. We also showed that inhibition of ERN1 signaling enzyme function significantly modified the response of most studied gene expressions to glucose deprivation condition. Thus, effect of glucose deprivation on the expression level of NR3C2, AHR, and SGK1 genes was significantly stronger in ERN1 knockdown U87 glioma cells since the expression of NNT gene was resistant to glucose deprivation condition. Moreover, the inhibition of ERN1 enzymatic activities in U87 glioma cells led to up-regulation of ARHGAP35 gene expression and significant down-regulation of the expression of SGK3 gene in response to glucose deprivation condition. Conclusions. Results of this study demonstrated that glucose deprivation did not change the expression level of NR3C1 gene but it significantly affected the expression of NR3C2, AHR, NRIP, SGK1, SGK3, and NNT genes in vector-transfected U87 glioma cells in gene specific manner and possibly contributed to the control of glioma growth since the expression of most studied genes in glucose deprivation condition was significantly dependent on the functional activity of IRE1 signaling enzyme.
- Published
- 2019
30. Hypoxic regulation of EDN1, EDNRA, EDNRB, and ECE1 gene expressions in ERN1 knockdown U87 glioma cells
- Author
-
Dmytro O. Minchenko, Myroslava Y. Sliusar, O O Riabovol, Yuliia M. Viletska, Daria O. Tsymbal, Yuliia O. Lahanovska, Borys H. Bezrodnyi, and Oleksandr H. Minchenko
- Subjects
0301 basic medicine ,microrna ,Endothelin converting enzyme 1 ,Endocrinology, Diabetes and Metabolism ,Endothelin-Converting Enzymes ,Protein Serine-Threonine Kinases ,Diseases of the endocrine glands. Clinical endocrinology ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,ece1 ,Cell Line, Tumor ,Glioma ,Endoribonucleases ,microRNA ,medicine ,Humans ,ednra ,ednrb ,education ,Gene ,neoplasms ,education.field_of_study ,Gene knockdown ,Endothelin-1 ,Brain Neoplasms ,hypoxia ,u87 glioma cells ,ern1 knockdown ,edn1 ,Receptor, Endothelin A ,medicine.disease ,RC648-665 ,Receptor, Endothelin B ,Cell Hypoxia ,Cell biology ,nervous system diseases ,Gene Expression Regulation, Neoplastic ,ERN1 ,030104 developmental biology ,Real-time polymerase chain reaction ,HIF1A ,Gene Knockdown Techniques ,030220 oncology & carcinogenesis ,Tumor Hypoxia ,mrna expression - Abstract
Objective. The aim of the present investigation was to study the effect of hypoxia on the expression of genes encoding endothelin-1 (EDN1) and its cognate receptors (EDNRA and EDNRB) as well as endothelin converting enzyme 1 (ECE1) in U87 glioma cells in response to inhibition of endoplasmic reticulum stress signaling mediated by ERN1/IRE1 (endoplasmic reticulum to nucleus signaling 1) for evaluation of their possible significance in the control of glioma growth through ERN1 and hypoxia. Methods. The expression level of EDN1, EDNRA, EDNRB, and ECE1 genes as well as micro-RNA miR-19, miR-96, and miR-206 was studied in control and ERN1 knockdown U87 glioma cells under hypoxia by quantitative polymerase chain reaction. Results. It was shown that the expression level of EDN1, EDNRA, EDNRB, and ECE1 genes was up-regulated in ERN1 knockdown glioma cells in comparison with the control glioma cells, being more significant for endothelin-1. We also observed down-regulation of microRNA miR-206, miR-96, and miR-19a, which have specific binding sites in mRNA EDN1, EDNRA, and EDNRB, correspondingly, and can participate in posttranscriptional regulation of these mRNA expressions. Furthermore, inhibition of ERN1 endoribonuclease lead to up-regulation of EDNRA and ECE1 gene expressions and down-regulation of the expression level of EDN1 and EDNRB genes in glioma cells. Thus, the expression of EDNRA and ECE1 genes is regulated by ERN1 endoribonuclease, but EDN1 and EDNRB genes preferentially by ERN1 protein kinase. We have also shown that hypoxia enhanced the expression of EDN1, EDNRA, and ECE1 genes and that knockdown of ERN1 signaling enzyme function significantly modified the response of all studied gene expressions to hypoxia. Thus, effect of hypoxia on the expression level of EDN1 and ECE1 genes was significantly or completely reduced in ERN1 knockdown glioma cells since the expression of EDNRA gene was down-regulated under hypoxia. Moreover, hypoxia is induced the expression of EDNRB gene in ERN1 knockdown glioma cells. Conclusions. Results of this investigation demonstrate that ERN1 knockdown significantly increased the expression of endothelin-1 and its receptors as well as ECE1 genes by different mechanisms and that all studied gene expressions were sensitive to hypoxia. It is possible that hypoxic regulation of the expression of these genes is a result of complex interaction of variable ERN1 related transcription and regulatory factors with HIF1A and possibly contributed to the control of glioma growth.
- Published
- 2019
31. Glucose deprivation affects the expression of genes encoding cAMP-activated protein kinase and related proteins in U87 glioma cells in ERN1 dependent manner
- Author
-
Oksana O. Ratushna
- Subjects
0301 basic medicine ,Endocrinology, Diabetes and Metabolism ,Protein Serine-Threonine Kinases ,Diseases of the endocrine glands. Clinical endocrinology ,03 medical and health sciences ,0302 clinical medicine ,Endocrinology ,Cell Line, Tumor ,Endoribonucleases ,Humans ,RNA, Messenger ,Protein kinase A ,neoplasms ,Gene knockdown ,biology ,u87 glioma cells ,Endoplasmic reticulum ,glucose deprivation ,Genes, Homeobox ,ern1 knockdown ,Transfection ,homeobox genes ,Glioma ,AKAP12 ,RC648-665 ,Cyclic AMP-Dependent Protein Kinases ,Ubiquitin ligase ,PRKACA ,Cell biology ,nervous system diseases ,ERN1 ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,Glucose ,030220 oncology & carcinogenesis ,biology.protein ,mrna expression - Abstract
Objective. The aim of this investigation was to study the expression of genes encoding cAMP-activated protein kinase catalytic and regulatory A subunits (PRKACA and PRKAR1A) and related proteins such as cAMP-dependent protein kinase inhibitors A and G (PKIA and PKIG), catalytic subunit A of protein phosphatase 3 (PPP3CA), A-kinase anchoring protein 12 (AKAP12), and praja ring finger ubiquitin ligase 2 (PJA2) in U87 glioma cells in response to glucose deprivation in both control U87 glioma cells and cells with ERN1 (endoplasmic reticulum to nucleus signaling 1) knockdown, the major pathway of the endoplasmic reticulum stress signaling, for evaluation of possible significance of glucose deprivation in ERN1 dependent regulation of glioma growth. Methods. The expression level of PRKA related genes was studied in control (transfected by vector) and ERN1 knockdown U87 glioma cells under glucose deprivation by real-time quantitative polymerase chain reaction. Results. It was shown that the expression level of PRKACA and PKIA genes was down-regulated in control glioma cells treated by glucose deprivation, but PJA2 gene was up-regulated. At the same time, the expression of four other genes (PRKAR1A, PKIG, AKAP12, and PPP3CA) was resistant to this experimental condition. Furthermore, ERN1 knockdown of glioma cells significantly modified the effect glucose deprivation on the expression almost all studied genes. Thus, treatment of glioma cells with inhibited ERN1 enzymatic activity by glucose deprivation lead to a more significant down-regulation of the expression level of PKIA and to suppression PRKAR1A gene expressions. Moreover, the ERN1 knockdown introduced up-regulation of PKIG and AKAP12 gene expressions in glioma cells treated by glucose deprivation and eliminated the sensitivity of PJA2 gene to this experimental condition. Conclusions. Results of this investigation demonstrated that ERN1 knockdown significantly modified the sensitivity of most studied PRKA related gene expressions to glucose deprivation and that these changes are a result of complex interactions of variable endoplasmic reticulum stress related and unrelated regulatory factors and contributed to the suppression of glioma cell proliferation and their possibly chemoresistance.
- Published
- 2021
32. Inhibitory effects of B-cell lymphoma 2 on the vasculogenic mimicry of hypoxic human glioma cells.
- Author
-
JIANWEN LI, YIQUAN KE, MIN HUANG, SHUYUN HUANG, and YIMING LIANG
- Subjects
- *
LYMPHOMAS , *B cells , *GLIOMAS , *WESTERN immunoblotting , *MESSENGER RNA - Abstract
The aim of this study was to investigate the mechanisms and effects of B-cell lymphoma 2 (Bcl-2) on the vasculogenic mimicry (VM) of human glioma cells. U87 cells were cultured under hypoxic conditions and then divided into four groups: Control, 3-(5-hydroxymethyl-2-furyl)-1-ben-zylindazole (YC-1), ABT-737 and YC-1 + ABT-737. These groups were treated with the corresponding simulators. The expression of hypoxia-inducible factor-1α (HIF-1α), matrix metalloproteinase (MMP)-2, MMP-14 and Bcl-2 in each group was determined using a reverse transcription-quantitative poly-merase chain reaction and western blot analysis. Compared with that in the control group, the mRNA and protein expression of MMP-2, MMP-14 and Bcl-2 in the YC-1 and ABT-737 groups was significantly reduced. The expression of HIF-la, however, was only significantly reduced in the YC-1 group (P<0.05). Compared with those in the YC-1 + ABT-737 group, the expression levels of the four proteins in the YC-1 and ABT-737 groups were not significantly different, with the exception of the expression of HIF-1α in the ABT-737 group, which was significantly enhanced (P<0.05). The mRNA expression levels of HIF-1α, MMP-2 and MMP-14 in the YC-1 group were significantly different from those in the ABT-737 group (P<0.01); however, no significant difference was observed in the expression of Bcl-2. In conclusion, Bcl-2 may be an important factor in the VM formation of human malignant glioma U87 cells under hypoxic conditions. Certain functions of Bcl-2 may be attributed to the HIF-1α-MMP-2-MMP-14-VM channel, whereas other functions may be independent of the channel. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
- View/download PDF
33. GLUTAMINE DEPRIVATION EFFECT ON DEK, TPD52, BRCA1, ADGRE5, LIF, GNPDA1, AND COL6A1 GENE EXPRESSIONS IN IRE1 KNOCKDOWN U87 GLIOMA CELLS
- Author
-
O. Y. Luzina, Y. A. Garmash, A. P. Kharkova, A.Y. Kuznetsova, O. H. Minchenko, S. V. Danilovskyi, and O. S. Hnatiuk
- Subjects
U87 glioma cells ,0301 basic medicine ,IRE1 knockdown ,ADGRE5 ,lcsh:Biotechnology ,Mrna expression ,COL6A1 ,GNPDA1 genes ,03 medical and health sciences ,lcsh:TP248.13-248.65 ,Glioma ,medicine ,glutamine deprivation ,U87 ,neoplasms ,Gene ,Gene knockdown ,Chemistry ,DEK ,mRNA expression ,BRCA1 ,medicine.disease ,nervous system diseases ,Cell biology ,Glutamine ,030104 developmental biology - Abstract
To study effect of glutamine deprivation on the expression of genes encoding the key proliferation associated factors on a relation to inhibition of inositol requiring enzyme-1 IRE1 in U87 glioma cells was the aim of the research. It was shown that glutamine withdrawal down-regulated the expression of DEK, BRCA1, LIF, and COL6A1 genes in control glioma cells (transfected by empty vector), up-regulated ADGRE5 gene expression, and did not significantly change the expression of TPD52 and GNPDA. Inhibition of ІRE1 signaling enzyme activity modified the effect of glutamine deprivation on the expression of TPD52, BRCA1, LIF, DEK, ADGRE5, and COL6A1 genes: introduces the effect of glutamine deprivation on TPD52 and GNPDA1, reduced – on COL6A1, and enhanced – on ADGRE5, DEK, and BRCA1 in U87 glioma cells. Therefore, glutamine deprivation affect the expression level of most studied genes in U87 glioma cells in relation to the functional activity of IRE1 signaling enzyme, which is responsible for control of cell proliferation and glioma growth.
- Published
- 2017
34. Enhanced induction of cell cycle arrest and apoptosis via the mitochondrial membrane potential disruption in human U87 malignant glioma cells by aloe emodin.
- Author
-
Ismail, Samhani, Haris, Khalilah, Abdul Ghani, Abdul Rahman Izaini, Abdullah, Jafri Malin, Johan, Muhammad Farid, and Mohamed Yusoff, Abdul Aziz
- Subjects
- *
ALOE , *APOPTOSIS , *EXPERIMENTAL design , *GLIOMAS , *MITOCHONDRIA , *RESEARCH funding , *T-test (Statistics) , *DATA analysis software , *DESCRIPTIVE statistics - Abstract
Aloe emodin, one of the active compounds found in Aloe vera leaves, plays an important role in the regulation of cell growth and death. It has been reported to promote the anti-cancer effects in various cancer cells by inducing apoptosis. However, the mechanism of inducing apoptosis by this agent is poorly understood in glioma cells. This research is to investigate the apoptosis and cell cycle arrest inducing by aloe emodin on U87 human malignant glioma cells. Aloe emodin showed a time- and dose-dependent inhibition of U87 cells proliferation and decreased the percentage of viable U87 cells via the induction of apoptosis. Characteristic morphological changes, such as the formation of apoptotic bodies, were observed with confocal microscope by Annexin V-FITC/PI staining, supporting our viability study and flow cytometry analysis results. Our data also demonstrated that aloe emodin arrested the cell cycle in the S phase and promoted the loss of mitochondrial membrane potential in U87 cells that indicated the early event of the mitochondria-induced apoptotic pathway. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
35. Comparison of metabolite profiles in U87 glioma cells and mesenchymal stem cells
- Author
-
Jürchott, Kathrin, Guo, Ke-Tai, Catchpole, Gareth, Feher, Kristen, Willmitzer, Lothar, Schichor, Christian, and Selbig, Joachim
- Subjects
- *
METABOLISM , *COMPARATIVE studies , *GLIOMAS , *MESENCHYMAL stem cells , *GAS chromatography/Mass spectrometry (GC-MS) , *GLYCOLYSIS , *CANCER cells , *SYSTEMS biology - Abstract
Abstract: Gas chromatography–mass spectrometry (GC–MS) profiles were generated from U87 glioma cells and human mesenchymal stem cells (hMSC). 37 metabolites representing glycolysis intermediates, TCA cycle metabolites, amino acids and lipids were selected for a detailed analysis. The concentrations of these metabolites were compared and Pearson correlation coefficients were used to calculate the relationship between pairs of metabolites. Metabolite profiles and correlation patterns differ significantly between the two cell lines. These profiles can be considered as a signature of the underlying biochemical system and provide snap-shots of the metabolism in mesenchymal stem cells and tumor cells. [Copyright &y& Elsevier]
- Published
- 2011
- Full Text
- View/download PDF
36. ATM-dependent ERK signaling via AKT in response to DNA double-strand breaks.
- Author
-
Khalil, Ashraf, Morgan, Rhiannon N., Adams, Bret R., Golding, Sarah E., Dever, Seth M., Rosenberg, Elizabeth, Povirk, Lawrence F., and Valerie, Kristoffer
- Published
- 2011
- Full Text
- View/download PDF
37. Expression of ubiquitin specific peptidase and ATG7 genes in U87 glioma cells upon glutamine deprivation
- Author
-
V. V. Telychko, Oleksandr H. Minchenko, О. O. Riabovol, О. O. Ratushna, Dmytro O. Minchenko, and O V Halkin
- Subjects
U87 glioma cells ,mRNA expression ,Biology ,medicine.disease ,Biochemistry ,lcsh:Biochemistry ,Glutamine ,USPs ,Ubiquitin ,Glioma ,medicine ,biology.protein ,lcsh:QD415-436 ,glutamine deprivation ,U87 ,ATG7 ,Gene ,IRE1 inhibition - Abstract
We have studied the effect of glutamine deprivation on the expression of genes encoding for ubiquitin specific peptidases (USP) and ubiquitin activating enzyme E1-like protein/autophagy related 7 (GSA7/ATG7) in U87 glioma cells in relation to inhibition of inositol requiring enzyme-1 (IRE1). It was shown that exposure of control glioma cells (transfected by empty vector) upon glutamine deprivation led to suppression of USP1 and ATG7 mRNA expression and up-regulated USP25 mRNA. At the same time, glutamine deprivation did not significantly change USP4, USP10, USP14, and USP22 gene expressions in these cells. Inhibition of ІRE1 signaling enzyme function in U87 glioma cells increased effect of glutamine deprivation on the expression of USP1 gene and introduced sensitivity of USP4 and USP14 genes to this condition. Therefore, glutamine deprivation affected the expression level of most studied genes in gene specific manner in relation to the functional activity of IRE1 enzyme, a central mediator of endoplasmic reticulum stress, which controls cell proliferation and tumor growth.
- Published
- 2017
38. Expression of tumor growth related genes in IRE1 knockdown U87 glioma cells: effect of hypoxia
- Author
-
Oleksandr H. Minchenko, O. Y. Luzina, I A Garmash, Dmytro O. Minchenko, O. S. Hnatiuk, and O O Ratushna
- Subjects
0301 basic medicine ,U87 glioma cells ,IRE1 knockdown ,COL6A1 ,Biology ,Biochemistry ,TPD52 ,lcsh:Biochemistry ,03 medical and health sciences ,Glioma ,medicine ,Tumor growth ,lcsh:QD415-436 ,U87 ,Gene ,Gene knockdown ,hypoxia ,mRNA expression ,Hypoxia (medical) ,medicine.disease ,BRCA1 ,stomatognathic diseases ,030104 developmental biology ,Cancer research ,HOMER3 ,medicine.symptom - Abstract
We have studied the effect of IRE1 signaling enzyme knockdown as well as hypoxia on the expression of genes encoding the important tumor growth related proteins (BRCA1, DEK, BCL2L1, COL6A1, TPD52, HOMER3, and GNPDA1) in U87 glioma cells. It was shown that the expression level of breast cancer 1 early onset (BRCA1) and tumor protein D52 (TPD52) mRNAs are strongly up-regulated in U87 glioma cells by down-regulation of IRE1 expression in comparison with the control cells. At the same time the expression level of collagen, type VI, alpha 1 (COL6A1), DEK oncogene (DEK), glucosamine-6-phosphate deaminase 1 (GNPDA1) and homer homolog 3 (HOMER3) was significantly down-regulated in glioma cells under these experimental conditions. It was also shown that hypoxia up-regulated the expression level of COL6A1 and TPD52 mRNAs and down-regulated – BRCA1, DEK, and GNPDA1 mRNAs in control glioma cells and that down-regulation of IRE1, which control cell proliferation and tumor growth, modified the effect of hypoxia on the expression of COL6A1, DEK, BCL2L1, HOMER3, and GNPDA1 genes. The present study demonstrated that hypoxia affected the expression of most studied genes in IRE1-dependent manner.
- Published
- 2017
39. Hypoxic regulation of the expression of genes encoded estrogen related proteins in U87 glioma cells: eff ect of IRE1 inhibition
- Author
-
Ratushna Oo, Minchenko Do, Minchenko Oh, and Riabovol Oo
- Subjects
0301 basic medicine ,Endocrinology, Diabetes and Metabolism ,Tripartite Motif Proteins ,Endocrinology ,Cation Transport Proteins ,Regulation of gene expression ,Reverse Transcriptase Polymerase Chain Reaction ,Kinase ,Nuclear Proteins ,Glioma ,Transfection ,Endoplasmic Reticulum Stress ,Neoplasm Proteins ,Nuclear Receptor Interacting Protein 1 ,Cell biology ,DNA-Binding Proteins ,Gene Expression Regulation, Neoplastic ,Receptors, Estrogen ,ebbp ,mrna expression ,Signal transduction ,Receptors, Progesterone ,pgrmc2 ,Signal Transduction ,medicine.medical_specialty ,Ubiquitin-Protein Ligases ,slc39a6 ,Protein Serine-Threonine Kinases ,Biology ,nrip1 ,Diseases of the endocrine glands. Clinical endocrinology ,Mitochondrial Proteins ,03 medical and health sciences ,Cell Line, Tumor ,Internal medicine ,Endoribonucleases ,medicine ,Humans ,neoplasms ,Adaptor Proteins, Signal Transducing ,Cell Proliferation ,ire1 inhibition ,hypoxia ,u87 glioma cells ,Endoplasmic reticulum ,Membrane Proteins ,RC648-665 ,medicine.disease ,nervous system diseases ,030104 developmental biology ,e2ig5 ,Cell culture ,Unfolded protein response ,Tumor Hypoxia ,esrra ,Glioblastoma ,Transcription Factors - Abstract
Objective. The aim of the present study was to examine the effect of inhibition of endoplasmic reticulum stress signaling, mediated by IRE1 (inositol requiring enzyme 1), which is a central mediator of the unfolded protein response on the expression of genes encoded estrogen related proteins (NRIP1/RIP140, TRIM16/EBBP, ESRRA/NR3B1, FAM162A/E2IG5, PGRMC2/PMBP, and SLC39A6/LIV-1) and their hypoxic regulation in U87 glioma cells for evaluation of their possible significance in the control of glioma cells proliferation. Methods. The expression of NRIP1, EBBP, ESRRA, E2IG5, PGRMC2, and SLC39A6 genes in U87 glioma cells, transfected by empty vector pcDNA3.1 (control) and cells without IRE1 signaling enzyme function (transfected by dnIRE1) upon hypoxia, was studied by a quantitative polymerase chain reaction. Results. Inhibition of both enzymatic activities (kinase and endoribonuclease) of IRE1 signaling enzyme function up-regulates the expression of EBBP, E2IG5, PGRMC2, and SLC39A6 genes is in U87 glioma cells in comparison with the control glioma cells, with more significant changes for E2IG5 and PGRMC2 genes. At the same time, the expression of NRIP1 and ESRRA genes is strongly down-regulated in glioma cells upon inhibition of IRE1. We also showed that hypoxia increases the expression of E2IG5, PGRMC2, and EBBP genes and decreases NRIP1 and ESRRA genes expression in control glioma cells. Furthermore, the inhibition of IRE1 in U87 glioma cells decreases the eff ect of hypoxia on the expression of E2IG5 and PGRMC2 genes, eliminates hypoxic regulation of NRIP1 gene, and enhances the sensitivity of ESRRA gene to hypoxic condition. Furthermore, the expression of SLC39A6 gene is resistant to hypoxia in both the glioma cells with and without IRE1 signaling enzyme function. Conclusions. Results of this investigation demonstrate that inhibition of IRE1 signaling enzyme function affects the expression of NRIP1, EBBP, ESRRA, E2IG5, PGRMC2, and SLC39A6 genes in U87 glioma cells in gene specific manner and these changes possibly contribute to the suppression of the cell proliferation. Most of these genes are regulated by hypoxia and preferentially through IRE1 signaling pathway of endoplasmic reticulum stress.
- Published
- 2017
40. Activation of the nuclear transcription factor κB (NFκB) and differential gene expression in U87 glioma cells after exposure to the cytoprotector amifostine
- Author
-
Kataoka, Yasushi, Murley, Jeffrey S., Khodarev, Nikolai N., Weichselbaum, Ralph R., and Grdina, David J.
- Subjects
- *
GENE expression , *GLIOMAS - Abstract
Purpose : Amifostine has been approved as a therapy to decrease the incidence of moderate-to-severe xerostomia in patients undergoing postoperative radiation treatment for head-and-neck cancer. As a reducing agent capable of participating in intracellular reductive/oxidative processes, it has the potential to affect redox-sensitive transcription factors and gene expression. Amifostine’s active free thiol WR-1065 was investigated to determine its effect on nuclear transcription factor κB (NFκB) activation and subsequent gene expression in U87 glioma cells.Methods and Materials : The human glioma cell line U87 was grown to confluency and then exposed to WR-1065 at a concentration of 40 μM for times ranging from 30 min to 24 h. Changes in cell cycle were monitored by flow cytometry. The effect of WR-1065 on NFκB activation was determined by a gel shift assay. Changes in gene expression as a function of time of exposure to WR-1065 were determined by Northern blot and the Atlas Human cDNA Expression Array (Clontech, Palo Alto, CA). Changes in gene expression using the Atlas Array were verified by reverse transcriptase-polymerase chain reaction (RT-PCR) with gene-specific primers.Results : Exposure of U87 cells to 40 μM WR-1065 resulted in a marked activation of NFκB between 30 min and 1 h after treatment. Expression of MnSOD, an NFκB-responsive gene, was enhanced by over 2-fold after 16 h of treatment and remained elevated at 24 h. During this period of time, no changes in cell cycle distribution were observed. To assess changes in the expression levels of NFκB-responsive genes as a function of WR-1065 exposure, cDNA arrays containing 49 genes identified as having DNA-binding motifs for NFκB were used. Only five genes were found to be significantly affected at 1, 4, and/or 16 h of treatment. GST-3 and c-myc were repressed up to 2- and 4-fold, respectively. The expression levels of IL-2Ra, RANTES, and c-myb, in contrast, were enhanced up to 14-, 3-, and 2-fold, respectively. The remaining genes having NFκB-responsive elements in their promoter regions were either not expressed (20 genes) or were not affected (24 genes) by exposure to WR-1065.Conclusions : The redox-sensitive transcription factor NFκB can be activated in U87 glioma cells by the active thiol form of the cytoprotector amifostine. Activation of NFκB by the antioxidant WR-1065 is accompanied by a reduced expression of the oncogene c-myc and an enhanced expression of the antioxidant gene MnSOD, a gene whose expression in tumor cells is relatively low, but when overexpressed has been correlated with a suppression of the malignant phenotype. Activation of NFκB by WR-1065, however, results in selective rather than global changes in the expression of genes containing NFκB-responsive elements. [Copyright &y& Elsevier]- Published
- 2002
- Full Text
- View/download PDF
41. Effect of hypoxia on the expression of genes encoding insulin-like growth factors and some related proteins in U87 glioma cells without IRE1 function
- Author
-
Kharkova Ap, L L Karbovskyi, Oleksandr H. Minchenko, Oleh V. Halkin, and Dmytro O. Minchenko
- Subjects
0301 basic medicine ,Endocrinology, Diabetes and Metabolism ,Protein Serine-Threonine Kinases ,Diseases of the endocrine glands. Clinical endocrinology ,Receptor, IGF Type 1 ,stc2 ,03 medical and health sciences ,Endocrinology ,Somatomedins ,Cell Line, Tumor ,Glioma ,Endoribonucleases ,medicine ,Humans ,Gene ,Glycoproteins ,Insulin-like growth factor 1 receptor ,ire1 inhibition ,Regulation of gene expression ,hypoxia ,u87 glioma cells ,Kinase ,igf1r ,Transfection ,RC648-665 ,medicine.disease ,Molecular biology ,Cell Hypoxia ,Cell biology ,Gene Expression Regulation, Neoplastic ,igf1 ,030104 developmental biology ,Insulin-Like Growth Factor Binding Protein 4 ,Cell culture ,igfbp4 ,Intercellular Signaling Peptides and Proteins ,mrna expression ,Signal transduction ,Signal Transduction - Abstract
Objective. The aim of the present study was to investigate the effect of hypoxia on the expression of genes encoding insulin-like growth factors (IGF1 and IGF2), their receptor (IGF1R), binding protein-4 (IGFBP4), and stanniocalcin 2 (STC2) in U87 glioma cells in relation to inhibition of endoplasmic reticulum stress signaling mediated by IRE1 (inositol requiring enzyme 1) for evaluation of their possible significance in the control of tumor growth. Methods. The expression of IGF1, IGF2, IGF1R, IGFBP4, and STC2 genes in U87 glioma cells transfected by empty vector pcDNA3.1 (control) and cells without IRE1 signaling enzyme function (transfected by dnIRE1) upon hypoxia was studied by qPCR. Results. The expression of IGF1 and IGF2 genes is down-regulated in glioma cells without IRE1 signaling enzyme function in comparison with the control cells. At the same time, the expression of IGF1R, IGFBP4, and STC2 genes was up-regulated in glioma cells upon inhibition of IRE1, with more significant changes for IGFBP4 and STC2 genes. We also showed that hypoxia does not change significantly the expression of IGF1, IGF2, and IGF1R genes but up-regulated IGFBP4 and STC2 genes expression in control glioma cells. Moreover, the inhibition of both enzymatic activities (kinase and endoribonuclease) of IRE1 in glioma cells does not change significantly the effect of hypoxia on the expression of IGF1, IGF1R, and IGFBP4 genes but introduces sensitivity of IGF2 gene to hypoxic condition. Thus, the expression of IGF2 gene is resistant to hypoxia only in control glioma cells and significantly down-regulated in cells without functional activity of IRE1 signaling enzyme, which is central mediator of the unfolded protein response and an important component of the tumor growth as well as metabolic diseases. Conclusions. Results of this study demonstrate that the expression of IGF1 and IGF1R genes is resistant to hypoxic condition both in control U87 glioma cells and cells without IRE1 signaling enzyme function. However, hypoxia significantly up-regulates the expression of IGFBP4 gene independently on the inhibition of IRE1 enzyme. These data show that proteins encoded by these genes are resistant to hypoxia except IGFBP4 and participate in the regulation of metabolic and proliferative processes through IRE1 signaling.
- Published
- 2016
42. GLUCOSE DEPRIVATION AFFECTS THE EXPRESSION OF LONP1 AND CATHEPSINS IN IRE1 KNOCKDOWN U87 GLIOMA CELLS
- Author
-
O O Ratushna, O. H. Minchenko, A Y Kuznetsova, O O Riabovo, D O Minchenko, and O V Halkin
- Subjects
LONP1/PRSS15 ,0301 basic medicine ,Cathepsin ,Gene knockdown ,Chemistry ,lcsh:Biotechnology ,glucose deprivation ,M RNA EXPRESSION,CTS,LONP1/PRSS15,IRE1 INHIBITION,GLUCOSE DEPRIVATION,U87 GLIOMA CELLS,ЕКСПРЕСіЯ МРНК,LONP1/ PRSS15,ПРИГНіЧЕННЯ IRE1,ДЕФіЦИТ ГЛЮКОЗИ,КЛіТИНИ ГЛіОМИ U87,ЭКСПРЕССИЯ МРНК,УГНЕТЕНИЕ IRE1,ДЕФИЦИТ ГЛЮКОЗЫ,КЛЕТКИ ГЛИОМЫ U87 ,mRNA expression ,medicine.disease ,U87 glioma cells ,03 medical and health sciences ,Glucose deprivation ,030104 developmental biology ,lcsh:TP248.13-248.65 ,Glioma ,Cancer research ,medicine ,U87 ,CTS ,IRE1 inhibition - Abstract
Метою роботи було вивчення впливу дефіциту глюкози на експресію генів, що кодують LONP1/PRSS15 та катепсини у клітинах гліоми лінії U87 за умов пригнічення IRE1 inositol requiring enzyme-1. Показано, що дефіцит глюкози посилював експресію генів CTSA, CTSB, CTSD, CTSK, CTSL, CTSO та LONP1, але не впливав на експресію генів CTSC, CTSF та CTSS у контрольних (трансфікованих порожнім вектором) клітинах гліоми. Пригнічення функції сигнального ензиму IRE1 у клітинах гліоми лінії U87 змінювало ефект дефіциту глюкози на експресію більшості досліджених генів: нівелювало ефект дефіциту глюкози на гени CTSA та CTSO, індукувало на гени CTSC та CTSS, зменшувало на ген CTSK і посилювало на ген CTSL. Таким чином, дефіцит глюкози змінював рівень експресії більшості досліджених генів залежно від функціональної активності ензиму IRE1, центрального медіатора стресу ендоплазматичного ретикулума, що контролює процеси проліферації та росту пухлин.Целью работы было изучение влияния дефицита глюкозы на экспрессию генов, которые кодируют LONP1/PRSS15 и катепсины в клетках глиомы линии U87 при угнетении 1IRE1 inositol requiring enzyme-1. Показано, что дефицит глюкозы усиливал экспрессию генов CTSA, CTSB, CTSD, CTSK, CTSL, CTSO и LONP1/PRSS15, но не влиял на экспрессию генов CTSC, CTSF и CTSS в контрольных (трансфецированных пустым вектором) клетках глиомы. Угнетение функции сигнального энзима IRE1 в клетках глиомы линии U87 изменяло эффект дефицита глюкозы на экспрессию большинства изученных генов: нивелировало эффект дефицита глюкозы на гены CTSA и CTSO, индуцировало на гены CTSC та CTSS, уменьшало на ген CTSK и усиливало на ген CTSL. Таким образом, дефицит глюкозы изменял уровень экспрессии большинства изученных генов в зависимости от функциональной активности энзима IRE1, центрального медиатора стресса эндоплазматического ретикулума, который контролирует процессы пролиферации и роста опухолей.To study the effect of glucose deprivation on the expression of genes encoding for LONP1/PRSS15 and cathepsins in U87 glioma cells in relation to inhibition of inositol requiring enzyme-1 (IRE1) was the aim of the research. It was shown that glucose deprivation up-regulated the expression of CTSA, CTSB, CTSD, CTSK, CTSL, CTSO, and LONP1 genes and did not change the expression of CTSC, CTSF, and CTSS genes in control glioma cells (transfected by empty vector). Inhibition of ІRE1 signaling enzyme function in U87 glioma cells modified effect of glucose deprivation on the expression of most studied genes: removed the effect of glucose deprivation on CTSA and CTSO genes, introduces on CTSC and CTSS genes, reduced on CTSK gene, and enhanced on CTSL gene. Therefore, glucose deprivation affect the expression level of most studied genes in relation to the functional activity of IRE1 enzyme, a central mediator of endoplasmic reticulum stress, which control cell proliferation and tumor growth.
- Published
- 2016
43. IRE1 KNOCKDOWN MODIFIES THE GLUTAMINE AND GLUCOSE DEPRIVATION EFFECT ON THE EXPRESSION OF NUCLEAR GENES ENCODING MITOCHONDRIAL PROTEINS IN U87 GLIOMA CELLS
- Author
-
O. H. Minchenko, O O Riabovol, D O Minchenko, Dariia O. Tsymbal, and O O Ratushna
- Subjects
0301 basic medicine ,Gene knockdown ,Nuclear gene ,Chemistry ,lcsh:Biotechnology ,glucose and glutamine deprivation, mitochondrial proteins, IRE1 inhibition, U87 glioma cells ,medicine.disease ,Molecular biology ,Cell biology ,Glutamine ,U87 glioma cells ,03 medical and health sciences ,Glucose deprivation ,030104 developmental biology ,mitochondrial proteins ,lcsh:TP248.13-248.65 ,Glioma ,medicine ,U87 ,Mitochondrial protein ,glucose and glutamine deprivations ,IRE1 inhibition - Abstract
We have studied the glucose and glutamine deprivation effect on the expression of nuclear genes encoding mitochondrial proteins in U87 glioma cells in relation to inhibition of inositol requiring enzyme-1 (IRE1). It was shown that glutamine deprivation down-regulated the expression of mitochondrial (NADP+)-dependent isocitrate dehydrogenase 2 (IDH2), malic enzyme 2 (ME2), mitochondrial aspartate aminotransferase (GOT2), and subunit B of succinate dehydrogenase (SDHB) genes in control glioma cells in gene specific manner. At the same time, the expression level of malate dehydrogenase 2 (MDH2) and subunit D of succinate dehydrogenase (SDHD) genes in these cells was not changed upon glutamine deprivation. It was also shown that inhibition of ІRE1 signaling enzyme function in U87 glioma cells modified the glutamine deprivation effect on the expression of all studied genes. Furthermore, the expression of the majority of studied genes was resistant to glucose deprivation, except IDH2 and SDHB genes, which expression levels were slightly down-regulated. Inhibition of IRE1 modified the effect of glucose deprivation on ME2, SDHB, SDHD, and GOT2 genes expression. Therefore, glucose and glutamine deprivation affected the expression level of the majority of nuclear genes encoding mitochondrial proteins in relation to the functional activity of IRE1 enzyme, which is a central mediator of endoplasmic reticulum stress and controls cell proliferation and tumor growth., Целью работы было изучить влияние дефицита глюкозы и глютамина на экспрессию ядерных генов, кодирующих митохондриальные протеины, в клетках глиомы линии U87 при угнетении inositol requiring enzyme-1 — IRE1. Показано, что дефицит глютамина снижает экспрессию генов митохондриальной NADP+зависимой изоцитратдегидрогеназы 2 — IDH2, малик энзима 2 — ME2, митохондриальной аспартатаминотрансферазы GOT2 и субъединицы В сукцинатдегидрогеназы — SDHB в контрольных трансфецированных вектором без вставки клетках глиомы геноспецифически. В то же время уровень экспрессии генов малатдегидрогеназы 2 –MDH2 и субъединицы D сукцинатдегидрогеназы в этих клетках при дефиците глютамина существенно не изменяется. Установлено также, что угнетение функции сигнального энзима IRE1 в клетках глиомы линии U87 модифицирует эффект дефицита глютамина на экспрессию всех изученных генов. Экспрессия большинства исследованных генов является резистентной к дефициту глюкозы, за исключением генов IDH2 и SDHB, для которых она несколько снижается. Угнетение IRE1 в клетках глиомы линии U87 модифицирует эффект дефицита глюкозы на экспрессию генов ME2, SDHB, SDHD и GOT2. Таким образом, дефицит глюкозы и глютамина изменяет уровень экспрессии большинства ядерных генов, кодирующих митохондриальные протеины, в зависимости от функциональной активности энзима IRE1, который является центральным медиатором стресса эндоплазматического ретикулума и контролирует процессы пролиферации и роста опухолей.
- Published
- 2016
44. Inhibitory effects of B-cell lymphoma 2 on the vasculogenic mimicry of hypoxic human glioma cells
- Author
-
Jianwen Li, Yiquan Ke, Yiming Liang, Min Huang, and Shuyun Huang
- Subjects
U87 glioma cells ,Cancer Research ,Pathology ,medicine.medical_specialty ,medicine.diagnostic_test ,Oncogene ,hypoxia ,Articles ,General Medicine ,Cell cycle ,Biology ,medicine.disease ,Molecular medicine ,Molecular biology ,Immunology and Microbiology (miscellaneous) ,Western blot ,Apoptosis ,Glioma ,medicine ,Vasculogenic mimicry ,B-cell lymphoma 2 ,B-cell lymphoma ,vasculogenic mimicry - Abstract
The aim of this study was to investigate the mechanisms and effects of B-cell lymphoma 2 (Bcl-2) on the vasculogenic mimicry (VM) of human glioma cells. U87 cells were cultured under hypoxic conditions and then divided into four groups: Control, 3-(5-hydroxymethyl-2-furyl)-1-benzylindazole (YC-1), ABT-737 and YC-1 + ABT-737. These groups were treated with the corresponding simulators. The expression of hypoxia-inducible factor-1α (HIF-1α), matrix metalloproteinase (MMP)-2, MMP-14 and Bcl-2 in each group was determined using a reverse transcription-quantitative polymerase chain reaction and western blot analysis. Compared with that in the control group, the mRNA and protein expression of MMP-2, MMP-14 and Bcl-2 in the YC-1 and ABT-737 groups was significantly reduced. The expression of HIF-1α, however, was only significantly reduced in the YC-1 group (P
- Published
- 2014
45. Inhibition of IRE1 signaling affects the expression of genes encoded glucocorticoid receptor and some related factors and their hypoxic regulation in U87 glioma cells
- Author
-
Dmytro O. Minchenko, O O Ratushna, O O Riabovol, Oleksandr H. Minchenko, and Dariia O. Tsymbal
- Subjects
0301 basic medicine ,sgk3 ,nr3c1 ,sgk1 ,Endocrinology, Diabetes and Metabolism ,ncoa2 ,Protein Serine-Threonine Kinases ,Transfection ,Diseases of the endocrine glands. Clinical endocrinology ,Gene Expression Regulation, Enzymologic ,arhgap35 ,03 medical and health sciences ,Endocrinology ,Glucocorticoid receptor ,Receptors, Glucocorticoid ,Glioma ,Cell Line, Tumor ,Endoribonucleases ,medicine ,Humans ,neoplasms ,ire1 inhibition ,Kinase ,hypoxia ,u87 glioma cells ,Brain Neoplasms ,Endoplasmic reticulum ,medicine.disease ,RC648-665 ,nnt ,Cell Hypoxia ,Cell biology ,nervous system diseases ,Gene Expression Regulation, Neoplastic ,030104 developmental biology ,Gene Knockdown Techniques ,SGK1 ,Unfolded protein response ,mrna expression ,Signal transduction ,Signal Transduction - Abstract
Objective. The aim of the present investigation was to examine the effect of inhibition of endoplasmic reticulum stress signaling, mediated by IRE1 (inositol requiring enzyme 1), which is a central mediator of the unfolded protein response on the expression of genes encoding glucocorticoid receptor (NR3C1) and some related proteins (SGK1, SGK3, NCOA1, NCOA2, ARHGAP35, NNT) and their hypoxic regulation in U87 glioma cells for evaluation of their possible significance in the control of the glioma growth. Methods. The expression of NR3C1,SGK1,SGK3, NCOA1, NCOA2, ARHGAP35, and NNT genes in U87 glioma cells, transfected by empty vector pcDNA3.1 (control) and cells without IRE1 signaling enzyme function (transfected by dnIRE1) upon hypoxia, was studied by quantitative polymerase chain reaction. Results. Inhibition of IRE1 signaling enzyme function up-regulates the expression of NR3C1, SGK1, NCOA1, NCOA2, ARHGAP35, and NNT genes in U87 glioma cells in comparison with the control glioma cells, with more significant changes for NR3C1, SGK1, and NNT genes. At the same time, the expression of SGK3 gene is strongly down-regulated in glioma cells upon inhibition of IRE1. We have also shown that hypoxia increases the expression of NR3C1, SGK1, NCOA2, ARHGAP35, and NNT genes but decreases SGK3 and NCOA1 genes expression in control glioma cells. Moreover, the inhibition of both enzymatic activities (kinase and endoribonuclease) of IRE1 in U87 glioma cells enhances the eff ect of hypoxia on the expression of SGK1, SGK3, and NNT genes, but decreases the sensitivity of NR3C1 gene to hypoxic condition. Furthermore, the expression of NCOA1 gene is resistant to hypoxia in control glioma cells, but NCOA2 and ARHGAP35 genes are resistant to this condition in glioma cells without functional activity of IRE1 signaling enzyme. Conclusions. Results of this investigation demonstrate that inhibition of IRE1 signaling enzyme function affects the expression of NR3C1, SGK1, SGK3, NCOA1, NCOA2, ARHGAP35, and NNT genes in U87 glioma cells in gene specific manner and that all these genes are regulated by hypoxia preferentially through IRE1 signaling pathway of the endoplasmic reticulum stress.
- Published
- 2016
46. IRE1 inhibition affects the expression of insulin-like growth factor binding protein genes and modifies its sensitivity to glucose deprivation in U87 glioma cells.
- Author
-
Minchenko DO, Kharkova AP, Tsymbal DO, Karbovskyi LL, and Minchenko OH
- Subjects
- Brain Neoplasms genetics, Brain Neoplasms pathology, CCN Intercellular Signaling Proteins genetics, CCN Intercellular Signaling Proteins metabolism, Cell Line, Tumor, Endoribonucleases genetics, Gene Expression Regulation, Neoplastic, Glioma genetics, Glioma pathology, Humans, Insulin-Like Growth Factor Binding Proteins genetics, Protein Serine-Threonine Kinases genetics, RNA, Messenger metabolism, Repressor Proteins genetics, Repressor Proteins metabolism, Signal Transduction, Transfection, Brain Neoplasms enzymology, Endoribonucleases deficiency, Glioma enzymology, Glucose deficiency, Insulin-Like Growth Factor Binding Proteins metabolism, Protein Serine-Threonine Kinases deficiency
- Abstract
Objective: The aim of the present study was to investigate the effect of inhibition of endoplasmic reticulum stress signaling mediated by IRE1/ERN1 (inositol-requiring enzyme 1/endoplasmic reticulum to nucleus signaling 1) on the expression of genes encoding different groups of insulin-like growth binding proteins (IGFBP6 and IGFBP7) and CCN family (IGFBP8/CTGF/CCN2, IGFBP9/NOV/CCN3, IGFBP10/CYR61/CCN1, WISP1/CCN4, and WISP2/CCN5) and its sensitivity to glucose deprivation in U87 glioma cells., Methods: The expression of IGFBP6, IGFBP7, IGFBP8, IGFBP9, IGFBP10, WISP1, and WISP2 genes was studied by qPCR in control U87 glioma cells (wild-type) and its subline with IRE1 signaling enzyme loss of function upon glucose deprivation., Results: The expression of IGFBP8, IGFBP9, and WISP2 genes was up-regulated in control glioma cells upon glucose deprivation with most significant changes for IGFBP9 gene. At the same time, the expression of IGFBP6, IGFBP10, and WISP1 genes was resistant to glucose deprivation in these glioma cells, but the IGFBP7 gene expression was down-regulated. The inhibition of both enzymatic activities (kinase and endoribonuclease) of IRE1 in glioma cells modified the sensitivity of most studied gene expressions to glucose deprivation condition: introduced sensitivity of IGFBP10 and WISP1 genes to glucose deprivation, enhanced the effect of this deprivation on IGFBP7 and IGFBP9 gene expressions, and reduced this effect on WISP2 gene and induced suppressive effect of glucose deprivation on the expression of IGFBP8 gene. Furthermore, the inhibition of IRE1 strongly affected the expression of all studied genes in glioma cells upon regular growing condition in gene specific manner: up-regulated the expression levels of IGFBP7, IGFBP8, IGFBP10, WISP1, and WISP2 genes and down-regulated the IGFBP6 and IGFBP9 genes., Conclusions: The data of this investigation demonstrate that the expression of IGFBP7, IGFBP8, IGFBP9, and WISP2 genes are sensitive to glucose deprivation in U87 glioma cells and that inhibition of IRE1 signaling enzyme function may significantly affect the expression of all studied genes in the presence of glucose as well as modify the effect of glucose deprivation on the expression of most studied genes. These data also show that proteins encoded by these genes may participate in the regulation of metabolic and proliferative processes via IGF/INS receptors and possibly other signaling pathways as well, via IRE1 signaling, which is a central mediator of the unfolded protein response and an important component of the tumor growth and metabolic diseases.
- Published
- 2015
- Full Text
- View/download PDF
47. Expression of insulin-like growth factor binding protein genes and its hypoxic regulation in U87 glioma cells depends on ERN1 mediated signaling pathway of endoplasmic reticulum stress.
- Author
-
Minchenko DO, Kharkova AP, Karbovskyi LL, and Minchenko OH
- Subjects
- Brain Neoplasms metabolism, Cell Hypoxia genetics, Cell Line, Tumor, Endoplasmic Reticulum Stress genetics, Endoribonucleases antagonists & inhibitors, Gene Expression Regulation, Neoplastic, Gene Knockdown Techniques, Glioma metabolism, Humans, Insulin-Like Growth Factor Binding Proteins metabolism, Protein Serine-Threonine Kinases antagonists & inhibitors, Signal Transduction genetics, Brain Neoplasms genetics, Endoplasmic Reticulum Stress physiology, Endoribonucleases physiology, Glioma genetics, Insulin-Like Growth Factor Binding Proteins genetics, Protein Serine-Threonine Kinases physiology
- Abstract
Objective: The aim of the present study was to examine the association between the expression of insulin-like growth binding protein-1 and -2 (IGFBP1 and IGFBP2), insulin-like growth factor 2 mRNA binding protein 3/KH domain containing protein over-expressed in cancer (IGF2BP3/KOC1), and HtrA serine peptidase 1/serine protease with IGF-binding domain (HTRA1/PRSS11) genes and function of endoplasmic reticulum stress signaling mediated by ERN1 (endoplasmic reticulum to nucleus signaling 1) as well as the regulation of these genes by hypoxia in U87glioma cells., Methods: The expression of IGFBP1, IGFBP2, IGF2BP3, and HTRA1 genes in U87 glioma cells and its subline with ERN1 signaling enzyme loss of function, were analyzed by qPCR. Cells underwent to hypoxia exposure (3% oxygen, 16 h)., Results: The blockade of both enzymatic activities (kinase and endoribonuclease) of ERN1 in glioma cells led to a significant down-regulation of the expression of IGFBP1, IGFBP2, and IGF2BP3 genes and strong up-regulation of HTRA1. At the same time, the inhibition of ERN1 endoribonuclease significantly increased the expression of IGFBP1, IGFBP2, and HTRA1 genes and did not affect the IGF2BP3 gene expression. Hypoxia up-regulated the expression of IGFBP1 and IGFBP2 genes in control glioma cells, with more significant changes in IGFBP1 gene. Furthermore, effect of hypoxia on these gene expressions was significantly lower in glioma cells without ERN1 signaling enzyme function., Conclusions: Results of this study demonstrate the dependence of insulin-like growth binding proteins as well as IGF2BP3 and HTRA1 gene expressions in U87 glioma cells on ERN1 signaling enzyme function and hypoxia, indicating its participation in the regulation of metabolic and proliferative processes via IGF/INS receptors, because endoplasmic reticulum stress is an important component of tumor growth and metabolic diseases.
- Published
- 2015
- Full Text
- View/download PDF
48. Inhibitory effects of B-cell lymphoma 2 on the vasculogenic mimicry of hypoxic human glioma cells.
- Author
-
Li J, Ke Y, Huang M, Huang S, and Liang Y
- Abstract
The aim of this study was to investigate the mechanisms and effects of B-cell lymphoma 2 (Bcl-2) on the vasculogenic mimicry (VM) of human glioma cells. U87 cells were cultured under hypoxic conditions and then divided into four groups: Control, 3-(5-hydroxymethyl-2-furyl)-1-benzylindazole (YC-1), ABT-737 and YC-1 + ABT-737. These groups were treated with the corresponding simulators. The expression of hypoxia-inducible factor-1α (HIF-1α), matrix metalloproteinase (MMP)-2, MMP-14 and Bcl-2 in each group was determined using a reverse transcription-quantitative polymerase chain reaction and western blot analysis. Compared with that in the control group, the mRNA and protein expression of MMP-2, MMP-14 and Bcl-2 in the YC-1 and ABT-737 groups was significantly reduced. The expression of HIF-1α, however, was only significantly reduced in the YC-1 group (P<0.05). Compared with those in the YC-1 + ABT-737 group, the expression levels of the four proteins in the YC-1 and ABT-737 groups were not significantly different, with the exception of the expression of HIF-1α in the ABT-737 group, which was significantly enhanced (P<0.05). The mRNA expression levels of HIF-1α, MMP-2 and MMP-14 in the YC-1 group were significantly different from those in the ABT-737 group (P<0.01); however, no significant difference was observed in the expression of Bcl-2. In conclusion, Bcl-2 may be an important factor in the VM formation of human malignant glioma U87 cells under hypoxic conditions. Certain functions of Bcl-2 may be attributed to the HIF-1α-MMP-2-MMP-14-VM channel, whereas other functions may be independent of the channel.
- Published
- 2015
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.