miR-219-5p对胰腺癌细胞增殖、凋亡和侵袭的影响及机制

摘要

目的 探讨miR-219-5p对胰腺癌细胞增殖、凋亡和侵袭的影响及机制.方法 采用实时荧光定量PCR技术(qPCR)检测miR-219-5p在胰腺癌细胞系Panc-1、AsPC-1及Bxpc-3和正常胰腺导管上皮细胞系HPDE6-C7中的表达.选取Panc-1细胞系,分成两组,对照组(NC组)转染阴性对照序列scramble,miR-219-5p组转染miR-219-5p mimics,MTT实验和流式细胞仪分析两组细胞增殖和凋亡情况,采用Transwell实验测定两组细胞侵袭能力,Western blotting法检测表皮生长因子受体(EGFR)和裂解型Caspase 3(Cleaved Caspase-3)蛋白表达.结果 miR-219-5p在胰腺癌细胞系Panc-1、AsPC-1及Bxpc-3中的相对表达量低于正常胰腺导管上皮细胞系HPDE6-C7(P均<0.01);miR-219-5p组在72、96及120 h 490 nm处的OD值低于NC组(P均<0.01),细胞凋亡率高于NC组(P均<0.01);miR-219-5p组、NC组细胞侵袭数分别为(75.3±6.9)、(172.5±10.5)个/视野,两组细胞侵袭数比较,P<0.01;miR-219-5p组、NC组EGFR相对表达量分别为0.31±0.09、1.0,两组比较,P<0.01;miR-219-5p组、NC组Cleaved Caspase 3蛋白相对表达量分别为2.7±0.15、1.0,两组比较,P<0.01.结论 miR-219-5p在胰腺癌细胞系中呈低表达,miR-219-5p过表达可抑制胰腺癌细胞增殖和侵袭,并诱导细胞凋亡;其机制可能与miR-219-5p下调EGFR蛋白和上调Cleaved Caspase-3蛋白表达有关.%Objective To investigate the effects of miRNA-219-5p on the proliferation,apoptosis,and invasion of pancreatic carcinoma cells and its mechanism. Methods Real-time fluorescence quantitative PCR (qPCR)was carried out to determine miR-219-5p expression in the pancreatic carcinoma cell lines Panc-1,AsPC-1,and Bxpc-3 and normal pancreatic ductal epithelial cell line HPDE6-C7. Further,Panc-1 cells were chosen and were divided into two groups:the normal control group (NC group)(transfection with miR-219-5p scramble)and miR-219-5p group (transfection with miR-219-5p mimics). The cell proliferation and apoptosis were detected by MTT assay and flow cytometry. The invasion ability was checked by transwell assay. The expression of epidermal growth factor receptor (EGFR)and Cleaved Caspase-3 pro-teins was examined by Western blotting. Results MiR-219-5p expression was lower in pancreatic carcinoma cell lines Panc-1,AsPC-1,and Bxpc-3,as compared with that in the pancreatic ductal epithelial cell line HPDE6-C7 (all P <0. 01). The OD490nm of the miR-219-5p group was significantly lower than that of the NC group at 72,96,and 120 h (all P < 0. 01);the apoptosis rate was higher (all P < 0. 01). The invasive cells were (75. 3 ± 6. 9)/ Hp in the miR-219-5p group,which was significantly lower than that [(172. 5 ± 10. 5)/ Hp]in the NC group (P < 0. 01). The expressive level of EGFR in the miR-219-5p group was significantly lower than NC group (0. 31 ± 0. 09 vs. 1. 0,P < 0. 01). The expres-sion of Cleaved Caspase-3 was up-regulated in the miR-219-5p group as compared with that of NC group (2. 7 ± 0. 15 vs.1. 0,P < 0. 01). Conclusions The miR-219-5p is lowly expressed in pancreatic carcinoma cell lines. The over-expression of miR-219-5p can inhibit the proliferation and invasion of pancreatic carcinoma cells,and induce the apoptosis,possibly though the down-regulated protein expression of EGFR and up-regulated protein expression of Cleaved Caspase-3.