Metformin Induces Growth Inhibition and Cell Cycle Arrest by Upregulating MicroRNA34a in Renal Cancer Cells
Wei Xie, Lei Wang, Halei Sheng, Jing Qiu, Di Zhang, Le Zhang, Fan Yang, Dahai Tang, Kebin Zhang
Central Laboratory, Xinqiao Hospital, Third Military Medical University, Chongqing, China (mainland)
Med Sci Monit 2017; 23:29-37
Metformin is a widely used biguanide drug for the treatment of type 2 diabetes. It has been revaluated as a potential anti-cancer drug with promising activity in various tumors. However, the precise mechanisms underlying the suppression of cancer cells by metformin remain not well understood.
MATERIAL AND METHODS: In this study, human renal cell carcinoma cell line ACHN was used to investigate the anti-proliferation effect of metformin. A cell counting kit-8 assay was used to detect the cell viability. The cell cycle distribution and apoptosis were analyzed by flow cytometry. The expression of cyclin D1 and p27KIP1 was detected by Western blot. The underlying mechanism involving miRNA34a was further investigated by quantitative RT-PCR and transfection with miRNA inhibitor specific for miRNA34a in ACHN, 769-P, and A498 cells.
RESULTS: Metformin could significantly inhibit the proliferation of ACHN cells in a dose- and time-dependent manner. In addition, the results showed that metformin induced G0/G1 phase arrest and delayed entry into S phase in ACHN cells. It was shown that metformin downregulates the expression of cyclin D1 and increases the p27KIP1 level. Furthermore, metformin increased ACHN cell death. Lastly, miRNA34a was found to be upregulated by metformin in ACHN, 769-P, and A498 cells. Subsequently, it was demonstrated that inhibition of miRNA34a could partially attenuate the suppressive effect of metformin on renal cancer cell proliferation.
CONCLUSIONS: The study data revealed that metformin induced cell growth inhibition and cell cycle arrest partially by upregulating miRNA34a in renal cancer cells.
Keywords: Apoptosis - drug effects, Antineoplastic Agents - pharmacology, Carcinoma, Renal Cell - pathology, Cell Cycle Checkpoints - drug effects, Cell Line, Tumor, Cell Proliferation - drug effects, Cell Survival - drug effects, Kidney Neoplasms - pathology