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目的:探索miR-219-5p对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞增殖、凋亡及侵袭的影响,并探讨其机制。方法:采用RT-PCR检测miR-219-5p在NSCLC细胞系H1299,A549,H1975及正常肺上皮细胞系BEAS-2B中的表达。将NSCLC细胞系H1299分成对照组和miR-219-5p组,用Lipofectamine 2000分别转染miR-219-5p scramble和miR-219-5p mimics,采用MTT法、流式细胞术及Transwell实验分别检测比较两组细胞增殖、凋亡及侵袭能力,Western印迹测定表皮生长因子受体(epidermal growth factor receptor,EGFR)及裂解型多聚腺苷二磷酸核糖聚合酶(poly ADP ribose polymerase,PARP)在两组细胞中的表达。结果:miR-219-5p在H1299,A549和H1975细胞系中的表达量均低于BEAS-2B,差异有统计学意义(P<0.05);MTT实验显示在48,72,96及120 h,miR-219-5p组OD490 nm值显著低于对照组,差异有统计学意义(P<0.05);miR-219-5p组细胞凋亡率显著高于对照组(13.33%±1.20%vs 3.43%±0.12%),差异有统计学意义(P<0.01);miR-219-5p组侵袭细胞数显著少于对照组(67.5±9.9 vs 189.5±16.7),差异有统计学意义(P<0.05);miR-219-5p组EGFR蛋白相对表达量为0.35±0.07,miR-219-5p组EGFR蛋白相对表达量显著低于对照组(1.0),差异有统计学意义(P<0.01);miR-219-5p组裂解型PARP蛋白相对表达量显著高于对照组(2.74±0.17 vs 1.0),差异有统计学意义(P<0.01)。结论:miR-219-5p可抑制NSCLC的细胞增殖和侵袭并促进其凋亡,其机制可能与下调EGFR及上调PARP的表达有关。
Objective: To explore the effect of miR-219-5p on proliferation, apoptosis and invasion of non-small cell lung cancer (NSCLC) cells and to explore its mechanism. Methods: The expression of miR-219-5p in NSCLC cell lines H1299, A549, H1975 and normal lung epithelial cell line BEAS-2B was detected by RT-PCR. The NSCLC cell line H1299 was divided into the control group and the miR-219-5p group. The miR-219-5p scramble and miR-219-5p mimics were transfected with Lipofectamine 2000 respectively. MTT assay, flow cytometry and Transwell assay were used to detect and compare The proliferation, apoptosis and invasion of the two groups of cells were examined by Western blotting. The expressions of epidermal growth factor receptor (EGFR) and poly ADP ribose polymerase (PARP) Expression in cells. Results: The expression level of miR-219-5p in H1299, A549 and H1975 cell lines was lower than that in BEAS-2B (P <0.05). MTT assay showed that the expression levels of miR-219-5p in H1299, The OD490 nm of miR-219-5p group was significantly lower than that of the control group (P <0.05), and the apoptosis rate of miR-219-5p group was significantly higher than that of the control group (13.33% ± 1.20% vs 3.43% ± 0.12%), the difference was statistically significant (P <0.01). The number of invasive cells in miR-219-5p group was significantly less than that in control group (67.5 ± 9.9 vs 189.5 ± 16.7, P <0.05) The relative expression level of EGFR protein in miR-219-5p group was 0.35 ± 0.07, the relative expression level of EGFR protein in miR-219-5p group was significantly lower than that in control group (P <0.01) The relative expression level of cleaved PARP protein in 219-5p group was significantly higher than that in control group (2.74 ± 0.17 vs 1.0), the difference was statistically significant (P <0.01). Conclusion: miR-219-5p can inhibit cell proliferation and invasion and promote apoptosis of NSCLC, which may be related to the down-regulation of EGFR and up-regulation of PARP expression.