miR-219-5p对非小细胞肺癌细胞增殖、凋亡与侵袭的影响及其机制
| 作者: |
1李磊,
1倪正义,
1汤中文,
1周密
1 武汉市医疗救治中心外一科，武汉 430000 |
| 通讯: |
周密
Email: mizhou768@126.com |
| DOI: | 10.3978/j.issn.2095-6959.2017.09.007 |
摘要
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。
关键词：
miR-219-5p
非小细胞肺癌
增殖
侵袭
凋亡
Effect of miR-219-5p on proliferation, apoptosis and invasion of non-small cell lung cancer and its mechanism
CorrespondingAuthor: ZHOU Mi Email: mizhou768@126.com
DOI: 10.3978/j.issn.2095-6959.2017.09.007
Abstract
Objective: To explore the role of miR-219-5p in the proliferation, apoptosis and invasion of non-small cell lung cancer (NSCLC) and study its mechanism. Methods: The expression of miR-219-5p in NSCLC cell lines H1299, A549, H1975 and BEAS-2B were evaluated, real-time quantitative PCR (RT-PCR) was determined. H1299 cell line was divided into a control group and a miR-219-5p group, which was transfected with miR-219-5p scramble and miR-219-5p mimic by Lipofectamine 2000, respectively. The proliferation, apoptosis and invasive of the two groups was measured by MTT assay, flow cytometry and Transwell assay. The expression level of EGFR and cleaved poly ADP ribose polymerase (PARP) protein was measured by Western blot. Results: The expression level of miR-219-5p in H1299, A549 and H1975 cell line was significantly lower than that in BEAS-2B (P<0.05). MTT showed that the value of OD490 nm of the miR-219-5p group was significantly lower than that in the control group in 48, 72, 96 and 120 h (P<0.05). The apoptosis rate of miR-219-5p was significantly higher than that in the control group (13.33%±1.20% vs 3.43%±0.12%; P<0.01). The number of invasive cells in the miR-219-5P group was significantly less than that in the control group (67.5±9.9 vs 189.5±16.7; P<0.01). The expression level of EGFR protein of miR-219-5p group was significantly lower than that in the control group (0.35±0.07 vs 1.0; P<0.01). The expression level of PARP protein of miR-219-5p group was significantly higher than that in the control group, (2.74±0.17 vs 1.0; P<0.01). Conclusion: miR-219-5p could restrain cell growth and invasion, and induce apoptosis in NSCLC cells, which may be possibly related with upregulated EGFR and downregulated PARP.
Keywords：
miR-219-5p
non-small cell lung cancer
proliferation
invasion
Apoptosis