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[Knockdown of AKT1 gene enhances the sensitivity of gastric cancer xenografts of nude mice to doxorubicin].
- Source :
-
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology [Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi] 2017 Oct; Vol. 33 (10), pp. 1354-1359. - Publication Year :
- 2017
-
Abstract
- Objective To investigate the effect of lentivirus-mediated shRNA targeting AKT1 gene on the sensitivity of gastric cancer xenografts to doxorubicin. Methods To establish the lentiviral vector of AKT1 RNA interference (RNAi), the vector of pGCSIL-shAKT1-GFP was infected into HEK293T cells, and meanwhile, the empty vector pGCSIL-shCON-GFP was assigned as a blank group, and then the viruses were harvested. BGC-823 gastric cancer cells were infected with the viruses. The protein expression of AKT1 was detected by Western blotting. The gastric cancer xenografts in nude mice were constructed using BGC-823 cells infected with the viruses, followed by the administration of doxorubicin. The size of tumor was evaluated and the growth curve of the tumor was drawn; HE staining was used to observe the pathological conditions of the xenografts; and the apoptosis of xenografts was examined by TUNEL assay. Results The recombinant plasmid of LV-shAKT1 was successfully constructed, and then transfected into HEK293T cells to produce high-titer lentivirus with a titer of 5×10 <superscript>8</superscript> TU/mL. The expression of AKT1 protein in gastric cancer BGC-823 cells was significantly down-regulated after successfully infected with the LV-shAKT1. Nude mice xenografts experiment showed that AKT1 gene silencing inhibited the growth of tumor xenografts, and also enhanced the inhibitory effect of doxorubicin on the growth of tumor xenografts. TUNEL staining showed that AKT1 gene silencing promoted the apoptosis of tumor xenograft cells, and the effect was more obvious when combined with doxorubicin. Conclusion AKT1 gene silencing can enhance the sensitivity of gastric cancer xenografts to doxorubicin through promoting cell apoptosis.
- Subjects :
- Animals
Antibiotics, Antineoplastic pharmacology
Apoptosis drug effects
Apoptosis genetics
Cell Line, Tumor
Cell Proliferation genetics
HEK293 Cells
Humans
Mice, Nude
Proto-Oncogene Proteins c-akt genetics
RNA Interference
Stomach Neoplasms genetics
Stomach Neoplasms pathology
Time Factors
Tumor Burden drug effects
Tumor Burden genetics
Cell Proliferation drug effects
Doxorubicin pharmacology
Proto-Oncogene Proteins c-akt metabolism
Stomach Neoplasms drug therapy
Xenograft Model Antitumor Assays methods
Subjects
Details
- Language :
- Chinese
- ISSN :
- 1007-8738
- Volume :
- 33
- Issue :
- 10
- Database :
- MEDLINE
- Journal :
- Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
- Publication Type :
- Academic Journal
- Accession number :
- 29169420