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This study explored the inhibitory effect of miR-223-3p on the proliferation of brain glioma cells and its potential mechanism in relation to growth arrest-specific transcripts (GAS5). We utilized the StarBase tool to predict the binding of GAS5 to miR-223-3p, further confirming the binding relationship between GAS5 and miR-223-3p through a dual luciferase reporter assay and RNA immunoprecipitation. In vitro cultured U87 brain glial cells were categorized into four groups: the control group, quercetin group, GAS5 inhibition group, and quercetin + si-GAS5 group. We analyzed and compared the impacts of quercetin on GAS5/miR-223-3p and pyroptotic proteins (NLRP3, ASC, caspase-1, GSDMD, and GSDME), as well as the proliferative capacity of U87 cells. StarBase's predictions indicated that GAS5 bound to miR-223-3p. This was corroborated by the dual luciferase reporter assay and RNA immunoprecipitation assay, which demonstrated a direct binding between GAS5 and miR -223-3p. Quercetin was observed to enhance GAS5 expression, inhibit the expression of miR-223-3p and pyroptotic proteins (NLRP3, ASC, caspase-1, GSDMD, and GSDME), and suppress the proliferation and invasion of U87 cells (P < 0.05). Downregulation of GAS5 expression correlated with an increase in the expression of miR-223-3p and pyroptotic proteins (NLRP3, ASC, caspase-1, GSDMD, and GSDME), promoting the proliferation and invasion of RC-4B/C cells (P < 0.05). Quercetin was able to reverse the increase in miR-223-3p expression and pyroptotic proteins (NLRP3, ASC, caspase-1, GSDMD, and GSDME) induced by the downregulation of GAS5, thereby inhibiting the proliferation and invasion of U87 cells (P < 0.05). In conclusion, quercetin might inhibit the proliferation of U87 brain glioma cells by promoting the expression of GAS5, which in turn competitively inhibited miR-223-3p and suppressed the pyroptotic signaling pathway. [ABSTRACT FROM AUTHOR]

Objective(s): Experimental studies reported that some plants in the genus of Moraea (Iridaceae family) show anticancer potential. This study aimed to evaluate the effects of Moraea sisyrinchium on U87 glioblastoma multiforme and HepG2 liver cancer cells.Materials and Methods: The cells were incubated for 24 hr with hydroalcoholic extract of the stem, flower, and bulb of M. sisyrinchium. Then, the cell proliferation (MTT) assay, cell cycle analysis (propidium iodide staining), cell migration test (scratch), Western blotting (Bax and Bcl-2 expression), and gelatin zymography (for matrix metalloproteinases, MMPs) were performed. Oxidative stress was evaluated by determining the levels of reactive oxygen species and lipid peroxidation. Angiogenesis was evaluated on chick embryo chorioallantoic membrane.Results: The extracts of the flower, stem, and bulb significantly decreased the proliferation of HepG2 and U87 cells. This effect was more for U87 than HepG2 and for the bulb and stem than the flower. In U87 cells, the bulb extract increased oxidative stress, cell cycle arrest, and the Bax/Bcl-2 ratio. Also, this extract suppressed the migration ability of HepG2 and U87 cells, which was associated with the inhibition of MMP2 activity. In addition, it significantly reduced the number and diameter of vessels in the chorioallantoic membrane. Liquid chromatography-mass spectrometry revealed the presence of xanthones (bellidifolin and mangiferin), flavonoids (quercetin and luteolin), isoflavones (iridin and tectorigenin), and phytosterols (e.g., stigmasterol) in the bulb.Conclusion: M. sisyrinchium bulb decreased the proliferation and survival of cancer cells by inducing oxidative stress. It also reduced the migration ability of the cells and inhibited angiogenesis.

Objective(s): Experimental studies reported that some plants in the genus of Moraea (Iridaceae family) show anticancer potential. This study aimed to evaluate the effects of Moraea sisyrinchium on U87 glioblastoma multiforme and HepG2 liver cancer cells. Materials and Methods: The cells were incubated for 24 hr with hydroalcoholic extract of the stem, flower, and bulb of M. sisyrinchium. Then, the cell proliferation (MTT) assay, cell cycle analysis (propidium iodide staining), cell migration test (scratch), Western blotting (Bax and Bcl-2 expression), and gelatin zymography (for matrix metalloproteinases, MMPs) were performed. Oxidative stress was evaluated by determining the levels of reactive oxygen species and lipid peroxidation. Angiogenesis was evaluated on chick embryo chorioallantoic membrane. Results: The extracts of the flower, stem, and bulb significantly decreased the proliferation of HepG2 and U87 cells. This effect was more for U87 than HepG2 and for the bulb and stem than the flower. In U87 cells, the bulb extract increased oxidative stress, cell cycle arrest, and the Bax/Bcl-2 ratio. Also, this extract suppressed the migration ability of HepG2 and U87 cells, which was associated with the inhibition of MMP2 activity. In addition, it significantly reduced the number and diameter of vessels in the chorioallantoic membrane. Liquid chromatography-mass spectrometry revealed the presence of xanthones (bellidifolin and mangiferin), flavonoids (quercetin and luteolin), isoflavones (iridin and tectorigenin), and phytosterols (e.g., stigmasterol) in the bulb. Conclusion: M. sisyrinchium bulb decreased the proliferation and survival of cancer cells by inducing oxidative stress. It also reduced the migration ability of the cells and inhibited angiogenesis. [ABSTRACT FROM AUTHOR]

Glycosaminoglycans (GAGs) such as heparan sulfate and chondroitin sulfate decorate all mammalian cell surfaces. These mucopolysaccharides act as coreceptors for extracellular ligands, regulating cell signaling, growth, proliferation, and adhesion. In glioblastoma, the most common type of primary malignant brain tumor, dysregulated GAG biosynthesis results in altered chain length, sulfation patterns, and the ratio of contributing monosaccharides. These events contribute to the loss of normal cellular function, initiating and sustaining malignant growth. Disruption of the aberrant cell surface GAGs with small molecule inhibitors of GAG biosynthetic enzymes is a potential therapeutic approach to blocking the rogue signaling and proliferation in glioma, including glioblastoma. Previously, 4-azido-xylose-α-UDP sugar inhibited both xylosyltransferase (XYLT-1) and β-1,4-galactosyltransferase-7 (β-GALT-7)-the first and second enzymes of GAG biosynthesis-when microinjected into a cell. In another study, 4-deoxy-4-fluoro-β-xylosides inhibited β-GALT-7 at 1 mM concentration in vitro. In this work, we seek to solve the enduring problem of drug delivery to human glioma cells at low concentrations. We developed a library of hydrophobic, presumed prodrugs 4-deoxy-4-fluoro-2,3-dibenzoyl-(α- or β-) xylosides and their corresponding hydrophilic inhibitors of XYLT-1 and β-GALT-7 enzymes. The prodrugs were designed to be activatable by carboxylesterase enzymes overexpressed in glioblastoma. Using a colorimetric MTT assay in human glioblastoma cell lines, we identified a prodrug-drug pair (4-nitrophenyl-α-xylosides) as lead drug candidates. The candidates arrest U251 cell growth at an IC50 = 380 nM (prodrug), 122 μM (drug), and U87 cells at IC50 = 10.57 μM (prodrug). Molecular docking studies were consistent with preferred binding of the α- versus β-nitro xyloside conformer to XYLT-1 and β-GALT-7 enzymes.

Glioblastoma (GBM) is an incurable brain tumor with a median survival of approximately 15 months despite an aggressive standard of care that includes surgery, chemotherapy, and ionizing radiation. Mouse models have advanced our understanding of GBM biology and the development of novel therapeutic strategies for GBM patients. However, model selection is crucial when testing developmental therapeutics, and each mouse model of GBM has unique advantages and disadvantages that can influence the validity and translatability of experimental results. To shed light on this process, we discuss the strengths and limitations of 3 types of mouse GBM models in this review: syngeneic models, genetically engineered mouse models, and xenograft models, including traditional xenograft cell lines and patient-derived xenograft models.

Glioblastoma multiforme (GBM) is a major aggressive primary brain tumor with dismal survival outcome and few therapeutic options. Although Temozolomide (TMZ) is a part of the standard therapy, over time, it can cause DNA damage leading to deleterious effects, necessitating the discovery of drugs with minimal side effects. To this end, we investigated the effect of cinnamaldehyde (CA), a highly purified, single ingredient from cinnamon, on the GBM cell lines U87 and U251 and the neuroglioma cell line H4. On observing similar impact on the viability in all the three cell lines, detailed studies were conducted with CA and its isomer/analog, trans-CA (TCA), and methoxy-CA (MCA) on U87 cells. The compounds exhibited equal potency when assessed at the cellular level in inhibiting U87 cells as well as at the molecular level, resulting in an increase in reactive oxygen species (ROS) and an increase in the apoptotic and multicaspase cell populations. To further characterize the key entities, protein profiling was performed with CA. The studies revealed differential regulation of entities that could be key to glioblastoma cell circuits such as downregulation of pyruvate kinase-PKM2, the key enzyme of the glycolytic pathway that is central to the Warburg effect. This allows for monitoring the levels of PKM2 after therapy using recently developed noninvasive technology employing PET [18F] DASA-23. Additionally, the observation of downregulation of phosphomevalonate kinase is significant as the brain tumor initiating cells (BTIC) are maintained by the metabolism occurring via the mevalonate pathway. Results from the current study, if translated in vivo, could provide additional efficacious treatment options for glioblastoma with minimal side effects. [ABSTRACT FROM AUTHOR]

The aim of this research was to discuss Hederagenin's antitumor effects on glioma by in vitro study. U251 and U87 cell lines were used as research target in our research. In the first step, the different Hed concentrations were treated to U251 and U87 cell lines, and the second step is Nur77 transfection in U251 and U87 with Hed treatment; measuring cell proliferation by MTT and EdU staining; evaluating cell invasion and migration abilities by transwell assay and relative gene and protein expressions by RT‐qPCR and WB assay. Compared with NC group, U251 and U87 cell proliferation were significantly depressed with cell apoptosis significantly increasing, and cell invasion and migration abilities were significantly inhibited in Hed‐treated groups (p <.05, respectively); however, with Nur77 transfection, the Hed's antitumor effects disappeared. Meanwhile, with Hed supplement, Nur77, PI3K, and AKT gene expressions were significantly downregulated (p <.05, respectively) in Hed‐treated groups; and Nur77, p‐PI3K, and p‐AKT protein expressions were significantly decreased (p <.05, respectively) in Hed‐treated groups. Hed had antitumor effects on glioma cell biological activities via Nur77/PI3K/AKT pathway in vitro study. [ABSTRACT FROM AUTHOR]

Cancer of the central nervous system (CNS) is ranked as the 19th most prevalent form of the disease in 2020. This study aims to identify candidate biomarkers and metabolic pathways affected by paclitaxel and etoposide, which serve as potential treatments for glioblastoma, and are linked to the pathogenesis of glioblastoma. We utilized an untargeted metabolomics approach using the highly sensitive ultra-high-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS) for identification. In this study, 92 and 94 metabolites in U87 and U373 cell lines were profiled, respectively. The produced metabolites were then analyzed utilizing t-tests, volcano plots, and enrichment analysis modules. Our analysis revealed distinct metabolites to be significantly dysregulated (nutriacholic acid, L-phenylalanine, L-arginine, guanosine, ADP, hypoxanthine, and guanine), and to a lesser extent, mevalonic acid in paclitaxel and/or etoposide treated cells. Furthermore, both urea and citric acid cycles, and metabolism of polyamines and amino acids (aspartate, arginine, and proline) were significantly enriched. These findings can be used to create a map that can be utilized to assess the antitumor effect of paclitaxel and/or etoposide within the studied cancer cells. [ABSTRACT FROM AUTHOR]

Vasculogenic mimicry (VM) is different from classical tumor angiogenesis and does not depend on endothelial cells. VM is closely related to the prognosis of various cancers. Canstatin was first identified as an endogenous angiogenesis inhibitor. In the present study, the inhibitory effect of canstatin on VM formation was evaluated. Human glioblastoma cell lines U87 and U251 were letivirally transduced to overexpress canstatin gene or GFP as control. In vitro assays showed that canstatin overexpression reduced the tube formation of U87 and U251 cells in Matrigel. A xenograft glioma model was created by subcutaneous injection of lentivirally modified U87 cells into nude mice. The results of in vivo experiments showed that canstatin gene introduction inhibited the growth of glioma xenografts. In tumor xenografts overexpressing canstatin, U87-mediated formation of VM-like structures and VM-related VEGF (vascular endothelial growth factor) expression were remarkably reduced. Canstatin overexpression also decreased the phosphorylation of Akt and reduced the expression of Survivin in vitro. In addition, HIF-1α production and MMP-2 secretion were decreased by canstatin overexpression. Therefore, these results suggested a protective role of canstatin during VM-like structure formation of glioma probably via inhibiting signaling pathways inducing vasculogenic mimicry.

Purpose: The aim of this study was to investigate the effects of Inula viscosa extract and Curcumin on the U87 (human astrocytoma cell line) treated with amyloid-beta (Aβ), which is the Alzheimer’s disease (AD) model cell line. Materials and Methods: Firstly, the cytotoxic potential of ınula and curcumin was investigated in the U87 cells by the colorimetric MTT (3-4,5-dimethyl-thiazolyl-2,5-diphenyltetrazolium bromide) assay. Then, the amount of Total Glutathione, Malondialdehyde (MDA), Glutathione reductase (GR) activities were investigated. ELISA test was used to examine the expression and activity of cleaved Bax and Bcl-2 proteins in the Inula viscosa and Curcumin treated U87 cell lines. Results: Inula viscosa and Curcumin treatment reduced cell death caused by amyloid-B in cells. It also reduced the oxidative stress caused by amyloid-B, while reducing the activation of the proapoptotic protein Bax, and Bcl-2. Conclusion: Our results suggest that inula viscosa may represent a new approach in the treatment of Alzheimer's.

Background & aim: The use of compounds extracted from natural plants due to less side effects can be a good alternative to chemical drugs in the treatment of deadly diseases such as cancer. In this study, the anticancer effect of Abu Jahl watermelon fruit extract on human glioblastoma cancer cells of U87 cell line was investigated. Therefore, the aim of this study was to determine and evaluate the effect of Citrullus colocynthis fruit extract on the expression of caspase 3 and caspase 8 genes in human glioblastoma cancer cell line.Using in vitro MTT assay and Real-time PCR, the cytotoxicity and cell inhibition mechanism of using Citrullus colocynthis on human glioblastoma U87 cell line was investigated. Methods: In this experimental study conducted in 2019 in the Faculty of Pharmacy and Pharmaceutical Sciences of Islamic Azad University, the fruit extract of the plant was prepared by extraction with 70% ethanol and to evaluate the toxicity of 104 cells of U87 cells were treated with different concentrations of this extract. These cells were treated as independent groups with specific concentrations of the extract at 24, 48 and 72 hours and were evaluated by MTT method. The determined IC50 concentration at 24 hours was used as the basal concentration for cell treatment to evaluate the amount of gene expression by Real-Time PCR. The collected data were analyzed using Kolmogorf-Smirnov statistical tests, one-way analysis of variance and Tukey. Results: The results obtained from MTT test showed that the growth of treated cells at concentrations higher than 1000 μg / ml of fruit extract was completely inhibited and the IC50 value calculated for 24, 48 and 72 hours of treatment was 1337.42, respectively. (P> 0.0001), 892.99 (p = 0.0012) and 387.49 (p> 0.0001) μg / ml of extract. Using IC50 value of Abu Jahl watermelon fruit extract for 24 hours increases the expression of caspase 3 and 8 gene by 3.052 (p = 0.0013) and 2.099 (p = 0.065), respectively. . Conclusion: The data revealed from this work determined that Citrullus colocynthis extract has concentration and time dependent toxicity effects on U87 cell line and the inhibition mechanism of this plant may be due to the induced Casp3 and Casp8 gene expression which leads to extrinsic apoptosis pathway. So, Citrullus colocynthis extract can be used to inhibit glioblastoma cells growth and may treat brain tumor.

BACKGROUND: Thymoquinone (TQ) is the active compound extracted from Nigella sativa which has been reported to possess various pharmacological attributes. This study shows that Thymoquinone (TQ) is an Alzheimer's disease (AD) model cell line on amyloid-beta (Aβ) -induced U87 (human astrocytoma cell line) in β-amyloid (Aβ)-induced in vitro Alzheimer's Disease (AD) model. AIM: We aimed to investigate the effects on antioxidant and apoptotic pathways. METHODS: In the study three groups were formed, the control group, the Aβ group, and the Aβ+TQ group obtained by adding TQ to the Aβ group. Firstly, the cytotoxic potential of TQ in U87 cells was investigated by the colorimetric MTT (3-4,5-dimethyl-thiazolyl-2,5 diphenyltetrazolium bromide) test. To determine the antioxidant status in the cell line treated with Thymoquinone, to examine the effects of superoxide dismutase (SOD) and catalase (CAT) activities, on apoptosis Caspase-3 protein levels were measured by ELISA method. RESULTS: When compared to the control group, the SOD and CAT levels were significantly decreased in the U87 cell line exposed to Aβ; Caspase-3 levels were found to increase significantly. However, application of TQ to the Aβ-U87 cell line significantly increased SOD and CAT levels; it was found that it decreased the caspase-3 level. CONCLUSION: In in vitro experiments, we determined that TQ has a protective effect by increasing antioxidant parameters in the amyloid beta-induced cell line (Fig. 4, Ref. 41). Text in PDF www.elis.sk [ABSTRACT FROM AUTHOR]

Glioblastoma is the most lethal malignancy of the brain and is resistant to conventional cancer treatments. Gene-therapy approaches like using tumor suppressor miRNAs are promising in the treatment of glioblastoma. They control the expression of oncogenes and influence tumor features and behaviors. Therefore, in the present study, it was predicted that miR-142 regulates oncogenic epidermal growth factor receptor (EGFR) signaling pathway via TargetScan and miRWalk online tools. Its differential expression level was reduced in glioblastoma according to the previous microarray results, and its predicted target genes were upregulated, as shown by the Expression Atlas. The miR-142 was overexpressed in U-87 glioblastoma cells via lentiviral transduction, and the way it influences proliferation and migration of cells was investigated through MTT assay and wound healing assay. Apoptosis rate was also measured via the Annexin V assay, and cell-cycle analysis was done. Then, real-time polymerase chain reaction (real-time PCR) and western blotting were performed to assess fold changes in mRNA and protein levels of the miR-142 predicted targets. Direct target genes of miR-142 were confirmed through a dual-luciferase reporter assay. The miR-142 significantly suppressed cell proliferation and migration and induced apoptosis and cell-cycle arrest in U-87 glioblastoma cells. This was accompanied by a decrease in expression of SHC adaptor protein 4 (SHC4), phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA), v-akt murine thymoma viral oncogene homolog 1 (AKT1), Kirsten rat sarcoma viral oncogene homolog (KRAS), and mitogen-activated protein kinase 8 (MAPK8) oncogenes at mRNA and protein levels in glioblastoma cells. Also, AKT1 was demonstrated as a direct target of miR-142. Overall, miR-424 acts as tumor suppressor miRNA in glioblastoma cells. [ABSTRACT FROM AUTHOR]

Cancer of the central nervous system (CNS) is ranked as the 19th most prevalent form of the disease in 2020. This study aims to identify candidate biomarkers and metabolic pathways affected by paclitaxel and etoposide, which serve as potential treatments for glioblastoma, and are linked to the pathogenesis of glioblastoma. We utilized an untargeted metabolomics approach using the highly sensitive ultra-high-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QTOF-MS) for identification. In this study, 92 and 94 metabolites in U87 and U373 cell lines were profiled, respectively. The produced metabolites were then analyzed utilizing t-tests, volcano plots, and enrichment analysis modules. Our analysis revealed distinct metabolites to be significantly dysregulated (nutriacholic acid, L-phenylalanine, L-arginine, guanosine, ADP, hypoxanthine, and guanine), and to a lesser extent, mevalonic acid in paclitaxel and/or etoposide treated cells. Furthermore, both urea and citric acid cycles, and metabolism of polyamines and amino acids (aspartate, arginine, and proline) were significantly enriched. These findings can be used to create a map that can be utilized to assess the antitumor effect of paclitaxel and/or etoposide within the studied cancer cells.

Objective To investigate the expression of phospho-c-Jun and MKK7 in gliomas and their correlation. Methods We collected and analyzed retrospectively 92 cases of gliomas, including 15 cases of diffuse astrocytoma, 5 cases of oligodendroglioma, 11 cases of anaplastic astrocytoma, 8 cases of anaplastic oligodendroglioma 53 cases of glioblastoma and 25 normal brain tissues adjacent to glioblastoma. The expression of c-Jun, phospho-c-Jun and MKK7 were detected by immunohistochemical staining. Glioma U87 cell line cultured in vitro was transfected with MKK4-siRNA, MKK7-siRNA and control siRNA for 48h, respectively. Western blot was performed to test the expression of c-Jun, phospho-c-Jun and MKK7. Results The expression of p-c-Jun and MKK7 in glioblastoma were significantly higher than those in other glioma and normal brain tissues adjacent to glioblastomas(P=0.000, P=0.000). The expression of MKK7 and phospho-c-Jun were positively correlated with WHO grading of gliomas (r＝0.494, P＝0.000; r=0.606, P=0.000). There was positive correlation between the expression of MKK7 and p-c-Jun (r=0.387, P=0.000). The knockdown of MKK7 suppressed c-Jun activities. Conclusion MKK7 promotes the development of glioblastoma by regulating the activity of c-Jun.

Purpose: The aim of this study was to investigate the effects of Inula viscosa extract and Curcumin on the U87 (human astrocytoma cell line) treated with amyloid-beta (Aβ), which is the Alzheimer’s disease (AD) model cell line. Materials and Methods: Firstly, the cytotoxic potential of ınula and curcumin was investigated in the U87 cells by the colorimetric MTT (3-4,5-dimethyl-thiazolyl-2,5- diphenyltetrazolium bromide) assay. Then, the amount of Total Glutathione, Malondialdehyde (MDA), Glutathione reductase (GR) activities were investigated. ELISA test was used to examine the expression and activity of cleaved Bax and Bcl-2 proteins in the Inula viscosa and Curcumin treated U87 cell lines. Results: Inula viscosa and Curcumin treatment reduced cell death caused by amyloid-B in cells. It also reduced the oxidative stress caused by amyloid-B, while reducing the activation of the proapoptotic protein Bax, and Bcl-2. Conclusion: Our results suggest that inula viscosa may represent a new approach in the treatment of Alzheimer's. [ABSTRACT FROM AUTHOR]

Background: Tankyrase1 (TNKS1), which often shows abnormal expression in many malignant tumor cells, plays an important role in tumor progression. In our previous study, we found that TNKS1 is also closely related to pathologic grade in human astrocytoma and its expression level is positively correlated with the Wnt/β-catenin pathway. This study is aimed to further elucidate the biological functions of TNKS1 as well as its relationship with the Wnt/β-catenin pathway. Methods: TNSK1 overexpression and knockdown vectors were constructed and transfected into glioblastoma cell lines U251 MG and U87, respectively. Viability, apoptosis, cell cycle and cell invasiveness in the treated cells were investigated. Results: In comparison with untreated cells, U251 and U87 cells overexpressing TNSK1 showed significantly increased cell viability and decreased apoptosis, while the TNKS1 knockdown U251 and U87 cells had reduced cell invasive ability and increased apoptosis, respectively. In addition, immunoprecipitation study showed that TNKS1 could be detected by β-catenin antibody after pull-down, indicating that TNKS1 directly interacts with β-catenin, further indicating that TNKS1 could be regarded as a positive regulator of the Wnt/β-catenin pathway in astrocytoma. Moreover, knockdown of TNKS1 in U251 and U87 cells also leads to suppressed Wnt/β-catenin signaling, and subsequent decrease of cell growth and proliferation, reduced invasion ability and increased apoptosis. Conclusion: Our findings suggest that TNKS1 might be a potential new therapeutic target for human astrocytoma in gene therapy. [ABSTRACT FROM AUTHOR]

Introduction: Glioblastoma multiforme (GBM) is a kind of malignant brain tumor prevalent in adults, with the characteristics well adapted to poorly immunogenic and hypoxic conditions. Effective treatment of GBM is impeded due to the high proliferation, migration and invasion of GBM cells. GBM cells migrate by degrading the extracellular matrix, so it is difficult to have GBM cells eradicated completely by surgery. This study aims to confirm that miR-431-5p could influence the proliferation, invasion and migration of human glioblastoma multiforme cells by targeting EPB41L1 (erythrocyte membrane protein band 4.1).Material and Methods: The expression levels of miR-431-5p and EPB41L1 were detected in GBM cells and tissues using qRT-PCR. Dual luciferase reporter gene assay and western blot were applied to confirm the targeting relationship between miR-431-5p and EPB41L1. GBM cell line U87 was used in MTT, flow cytometry, Transwell, and wound healing assays to determine cell proliferation, migration and invasion.Results: MiR-431-5p was overexpressed in GBM tissues while EPB41L1 was under-expressed. The results of dual luciferase reporter gene assay and western blot demonstrated that miR-431-5p could target EPB41L1 and suppress its expression. Down-regulating the expression of miR-431-5p or up-regulating the expression of EPB41L1 could inhibit the proliferation, invasion and migration but promote the apoptosis of GBM cells.Conclusions: MiR-431-5p facilitated the progression of GBM by inhibiting EPB41L1 expression. [ABSTRACT FROM AUTHOR]

Fatty acid-binding proteins (FABPs) bind and internalize long-chain fatty acids, controlling lipid dynamics. Recent studies have proposed the involvement of FABPs, particularly FABP7, in lipid droplet (LD) formation in glioma, but the physiological significance of LDs is poorly understood. In this study, we sought to examine the role of FABP7 in primary mouse astrocytes, focusing on its protective effect against reactive oxygen species (ROS) stress. In FABP7 knockout (KO) astrocytes, ROS induction significantly decreased LD accumulation, elevated ROS toxicity, and impaired thioredoxin (TRX) but not peroxiredoxin 1 (PRX1) signalling compared to ROS induction in wild-type astrocytes. Consequently, activation of apoptosis signalling molecules, including p38 mitogen-activated protein kinase (MAPK) and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK), and increased expression of cleaved caspase 3 were observed in FABP7 KO astrocytes under ROS stress. N-acetyl L-cysteine (NAC) application successfully rescued the ROS toxicity in FABP7 KO astrocytes. Furthermore, FABP7 overexpression in U87 human glioma cell line revealed higher LD accumulation and higher antioxidant defence enzyme (TRX, TRX reductase 1 [TRXRD1]) expression than mock transfection and protected against apoptosis signalling (p38 MAPK, SAPK/JNK and cleaved caspase 3) activation. Taken together, these data suggest that FABP7 protects astrocytes from ROS toxicity through LD formation, providing new insights linking FABP7, lipid homeostasis, and neuropsychiatric/neurodegenerative disorders, including Alzheimer's disease and schizophrenia. [ABSTRACT FROM AUTHOR]

• The glioblastoma cell line U87 can be differentiated into cholinergic neuron characteristics. • Sodium butyrate can induce the U87 cells differentiation into cells with cholinergic neuron characteristics. • Akt participates in the process of U87 cell differentiation into cholinergic neuron characteristics. To facilitate research methodologies for investigating the role of cholinergic nerves in many diseases, establishing an in vitro cholinergic neuron model is necessary. In this study, we investigated whether human glioblastoma U87 cells could be differentiated into cholinergic neurons in vitro. Sodium butyrate was used as the differentiation agent. The differentiated cells established by inducing U87 cells with sodium butyrate were named D-U87 cells. Immunofluorescence was used to label the neuronal markers MAP2, NF-M, and ChAT and the glial marker GFAP in D-U87 cells. Flow cytometry was used to measure cell cycle distribution in D-U87 cells. PCR, protein chip, and western blot assays were used to measure the expression levels of muscarinic cholinergic receptor 1 (M1), M4, ChAT, SYP and Akt. ELISA was used to measure neurotransmitter levels. As a result, we found that sodium butyrate induced U87 cell differentiation into cells with neuronal characteristics and increased not only the expression levels of the cholinergic neuron-related proteins M1, M4, ChAT and SYP in D-U87 cells but also the acetylcholine neurotransmitters in D-U87 cells. Moreover, the Akt protein expression in D-U87 cells was increased compared with that in U87 cells. Finally, we found that M1, M4, ChAT and SYP protein expression and acetylcholine secretion levels were significantly decreased in D-U87 cells after treatment with the Akt inhibitor MK-2206. These results demonstrate that D-U87 cells exhibit cholinergic neuron characteristics and that sodium butyrate induced U87 cell differentiation into cholinergic neuron partially through Akt signaling. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the effect of LncRNA ZEB1-AS1 on the proliferation, invasion and apoptosis of human glioma U87 cells. U87 glioma cells were divided into three groups. The Si group was transfected with LncRNA ZEB1-AS1 specific SiRNA. The NC group was transfected with non-specific scramble siRNA, and untransfected glioma cells were used as the blank group. After 48 h of transfection, the proliferation of U87 cells was detected by MTT assay, apoptosis of U87 cells was detected by flow cytometry, and Transwell invasion assay was used to detect cell invasion. The expression of LncZEB1-AS1 in Si group was significantly lower than that in the NC and blank groups (P<0.01). There was no statistical difference in the OD 490 between the three groups at 24 h (P>0.05). At 48 h, the Si group was significantly lower than the NC group and the blank group (P<0.01). After 48 h, the three groups showed a gradually increasing trend, but at all the time points, the Si group was always lower than the NC and blank groups (P<0.01). The OD values of the blank and NC groups were significantly higher than the same group at the previous time point (P<0.01). The OD values of Si group at 48 and 96 h were significantly higher than those at the previous time point (P <0.05). Although there was an upward trend between 72 and 48 h, the difference was not significant (P>0.05). Flow cytometry detected apoptosis in each group and found that the apoptosis rate in the Si group was significantly higher than that in the NC and blank groups (P<0.01). Inhibition of LncRNA ZEB1-AS1 can inhibit the proliferation and invasion of glioma U87 cells and promote apoptosis. LncRNA ZEB1-AS1 is expected to become a new target for the treatment of glioma. [ABSTRACT FROM AUTHOR]

Summary: Aim: Glioma, with fast growth and progression features, is the most common and aggressive tumor in the central nervous system and is essentially incurable. This study is aimed at inducing neuronal differentiation to suppress glioma cell growth with a single transcription factor. Methods: Overexpression of transcription factor SRY (sex determining region Y)‐box 11 (SOX11) and Zic family member 1 (ZIC1) was, respectively, performed in glioma cells with lentivirus infection. CRISPR/Cas9 technology was used to knock out ZIC1 in U87 cells, and knockout efficiency was identified by Western blotting and Sanger sequencing. Cell cycle and apoptosis were detected by flow cytometry. The downstream targets of SOX11 were analyzed by Affymetrix GeneChip microarrays. qRT‐PCR and immunofluorescence technique were used to verify gene targets of genetically modified U87 cells. All the cells were imaged by a fluorescence microscope. Gene expression correlation analysis and overall survival analysis based on TCGA dataset are performed by GEPIA. Results: We induced glioma cells into neuron‐like cells to suppress cell growth using a single transcription factor, SOX11 or ZIC1. Besides, we proved that there is a strong correlation between SOX11 and ZIC1. Our study revealed that SOX11 upregulates ZIC1 expression by binding with ZIC1 promoter, and ZIC1 partially mediates SOX11‐induced neuronal differentiation in U87 cells. However, SOX11 expression is not regulated by ZIC1. Moreover, high MAP2 expression means better overall survival in TCGA lower grade glioma. Conclusion: This study revealed that glioma cells can be reprogrammed into neuron‐like cells using a single factor ZIC1, which may be a potential tumor suppressor gene for gliomas treatment. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Magnetic NiFe 2 O 4 rod-like platelet were prepared in the presence of Utrica as a novel green precursor. • Prepared nanoparticles indicated more cytotoxicity effects after 48 h of frequency magnetic field exposure. • Therapeutic properties of low frequency of magnetic field by MNPs on neural cells was indicated. Abstract Despite of great attention concerned on Ni ferrite nanostructures in bioapplications, little is known about the toxicity of these NPs at the cellular and molecular levels. U87 (human primary glioblastoma) and SH-SY5Y (human neuroblastoma) cells treated with various concentration of well-characterized magnetic nickel ferrite nanoparticles, exposed to frequency magnetic field (FMF) and their response was studied. Ferromagnetic nanocrystalline nickel ferrite (NiFe 2 O 4) powder that characterized by XRD, TEM, SEM, FT-IR, nanosizer, and VSM techniques were prepared by a hydrothermal method in the presence of Urtica plant extract as a green precursor that acts both as reducing and capping agent. Owing to the exceptional properties of green alkalinized agent such as minor toxicity, higher biodegradability, high active surface and environment compatibility, we used the green alkalinized agent (Utrica) to prepared nanostructures for the first time. According to the obtained results, the FMF exposure caused an increase in cell death in neural cell types 48 h after treatment. MNPs indicated dose-dependent cytotoxicity but the amount of cell death per cell in the absence of MFM for SH-SY5Y cells was more than in U87 cells. On the other hand, cell death induced by FMF exposure was observed specifically in SH-SY5Y cells. Nevertheless, it is essential to perform more investigations to find the exact related mechanisms. Imatinib showed dose-dependent antiproliferative effects in all three prostate cancer cell lines. [ABSTRACT FROM AUTHOR]

Cellular NAD(P)H-dependent oxidoreductase activity with artificial dyes (NAD(P)H-OR) is an indicator of viability, as the cellular redox state is important for biosynthesis and antioxidant defense. However, high NAD(P)H due to impaired mitochondrial oxidation, known as reductive stress, should increase NAD(P)H-OR yet perturb viability. To better understand this complex behavior, we assayed NAD(P)H-OR with resazurin (Alamar Blue) in glioblastoma cell lines U87 and T98G, treated with inhibitors of central metabolism, oxythiamin, and phosphonate analogs of 2-oxo acids. Targeting the thiamin diphosphate (ThDP)-dependent enzymes, the inhibitors are known to decrease the NAD(P)H production in the pentose phosphate shuttle and/or upon mitochondrial oxidation of 2-oxo acids. Nevertheless, the inhibitors elevated NAD(P)H-OR with resazurin in a time- and concentration-dependent manner, suggesting impaired NAD(P)H oxidation rather than increased viability. In particular, inhibition of the ThDP-dependent enzymes affects metabolism of malate, which mediates mitochondrial oxidation of cytosolic NAD(P)H. We showed that oxythiamin not only inhibited mitochondrial 2-oxo acid dehydrogenases, but also induced cell-specific changes in glutamate and malate dehydrogenases and/or malic enzyme. As a result, inhibition of the 2-oxo acid dehydrogenases compromises mitochondrial metabolism, with the dysregulated electron fluxes leading to increases in cellular NAD(P)H-OR. Perturbed mitochondrial oxidation of NAD(P)H may thus complicate the NAD(P)H-based viability assay.

Novel treatment options are urgently required for patients with glioma who are not effectively treated through standard therapy. Human sodium iodide symporter (hNIS) is a molecular target of certain tumors types. Compared with 131I, 188Re possesses a higher energy and shorter half‑life; therefore, the effects of 188Re and 131I were compared in hNIS‑mediated gene imaging and therapy in the present study. Recombinant human brain glioma cell line U87 was transfected with a recombinant lentiviral vector containing hNIS (U87‑hNIS). U87‑0 cell line transfected with blank lentivirus was prepared as a control. In vitro, the 188Re and 131I uptake of U87‑hNIS cells were 21.3‑times and 25.9‑times that of the control groups, however the excretion rate of the two nuclides was very rapid, and the half‑life was only ~4 min. Sodium perchlorate inhibited hNIS‑mediated 188Re and 131I uptake to levels observed in the control groups. 188Re and 131I were able to kill U87‑hNIS cells selectively, with a survival of only 21.6 and 36.2%, respectively. U87‑hNIS nude mice appeared to accumulate 188Re, with a ratio of radioactivity counts between tumor and non‑tumor sites of ~13.5 compared with 10.3 of 131I 1 h after radionuclide injection. In contrast with in vitro studies, U87‑hNIS cells demonstrated a notable increase in 188Re retention in vivo, even 24 h after 188Re injection. U87‑hNIS cells also exhibited increased 131I retention in vivo; however, as the time increased, 131I was rapidly released with the tumor no longer able to be imaged 24 h after 131I injection. Following treatment, U87‑hNIS tumors experienced a volume reduction of 24.1%, whereas U87‑0 cells demonstrated an increase of 28.8%. 188Re and 131I were revealed to be effective at decreasing tumor volume compared with the control. However, 188Re was significantly more potent compared with 131I (P<0.01). The present study indicated that the U87‑hNIS cell line is sufficient to induce specific 188Re and 131I uptake, which may kill cells in vitro and in vivo. 188Re exhibited an increased retention time in vivo compared with 131I, which facilitates the imaging and therapy of U87‑hNIS tumors. [ABSTRACT FROM AUTHOR]

Glioblastoma (GBM) remains as the most common and aggressive brain tumor. The survival of GBM has been linked to the aberrant activation of swelling-induced chloride current ICl,swell. In this study, we investigated the effects of ICl,swell on cell viability, proliferation, and migration in the human GBM cell lines, U251 and U87, using a combination of patch clamp electrophysiology, MTT, colony formation, wound healing assays and Western immunoblotting. First, we showed that the specific inhibitor of ICl,swell, DCPIB, potently reduced the ICl,swell in U87 cells. Next, in both U87 and U251 cells, we found that DCPIB reduced GBM viability, proliferation, colony formation, migration, and invasion. In addition, our Western immunoblot assay showed that DCPIB-treated U251 cells had a reduction in JAK2, STAT3, and Akt phosphorylation, thus, suggesting that DCPIB potentially suppresses GBM functions through inhibition of the JAK2/STAT3 and PI3K/Akt signaling pathways. Therefore, the ICl,swell may be a potential drug target for GBM. [ABSTRACT FROM AUTHOR]