Effect of microRNA-216 on the proliferation and apoptosis of pancreatic cancer cells and the drug resistance to gemcitabine

XING Xin; GUO Yanwei; WANG Lijuan; ZHANG Hongqiao; ZHANG Jian; ZHENG Xiaoke; WANG Yanan

doi:10.3969/j.issn.1009-0460.2018.01.004

Abstract：Objective To explore the effect of microRNA-216 (miR-216) on the proliferation and apoptosis of pancreatic cancer cells and the drug resistance to gemcitabine. Methods The real-time quantitative PCR (QPCR) method was used to detect the miR-216 level in the gemcitabine resistant cell line BxPC-3 and the non-drug-resistant cell line CFPAC-1 of pancreatic cancer. The miR-216 mimics (mimics group) and inhibitor (inhibitor group) were transfected to BxPC-3 cells by Lipofectamine 2000 liposome method. BxPC-3 cells transfected with liposomes were used as the control group. QPCR was used to detect the level of miR-216 at 48 h after transfection. The CCK-8 method was used to detect the absorbance of each group at 24, 48, and 72 h after transfection to evaluate the proliferation rate. Annexin V-FITC/PI double staining was used to detect the apoptotic rates of each group at 48 h after transfection. The CCK-8 method was used to detect the half inhibitory concentration (IC50) of gemcitabine. The target gene of miR-216 was predicted by bioinformatics database miRBase and Gene Oncology (GO) function was annotated by cytoscape 3.5.1 and its plug-in Cl-uGO. Results The results of QPCR detection showed that the level of miR-216 in BxPC-3 cells was 3.010±0.901, higher than 1.049±0.074 of CFPAC-1 cells (P<0.05). The expression levels of miR-216 in the control group, mimics group and inhibitor group were1.130±0.145, 4.843±0.782 and 0.250±0.145(P<0.05). Compared with the control group, the proliferative rates increased and apoptotic rates decreased in mimics group while the proliferative rates decreased and apoptotic rates increased in inhibitor group (P<0.05). The IC50 of gemcitabine were (2.134±0.591) μg/ml, (4.518±0. 862) μg/ml and (0.481 ±0.073) μg/ml in the control group, mimics group and inhibitor group. The resistance of BxPC-3 cells to gemcitabine decreased after transfection of miR-216-inhibitor, and the resistance to gemcitabine increased after transfection of miR-216-mimics (P<0.05). There were 138 predicted target genes of miR-216. GO functions are mainly enriched in proliferation, apoptosis, invasion and migration. Conclusion The expression of miR-216 is up-regulated in pancreatic cancer drug resistant cells and can induce the proliferation of cancer cells, suggesting that it can play a similar role in promoting tumor genes and participate in the process of gemcitabine resistance, and it may become a target for early diagnosis and new biological treatment of pancreatic cancer.