上调microRNA-214表达对肺癌细胞凋亡及耐药性的影响-东南大学学报医学版

上调microRNA-214表达对肺癌细胞凋亡及耐药性的影响

单位：武汉市普仁医院呼吸内科, 湖北武汉 430081

分类号：R734.2

出版年·卷·期（页码）：2017·36·第五期（811-815）

摘要：

目的：探究microRNA-214过表达是否可以通过调节凋亡过程影响肺癌细胞的多药耐药性。方法：选择2种不同的人小细胞肺癌细胞株和人非小细胞肺癌细胞株作为实验组，分别是H69细胞实验组和H446细胞实验组；A549细胞实验组和H1299细胞组；选择正常人肺上皮细胞（BEAs-2B）作为对照组。Real-time PCR法进行检测肺癌细胞株内miR-214的表达量。上调miR-214表达后给予不同的抗肿瘤药物刺激，并MTT比色法检测肺癌细胞株对药物的敏感性；CCK-8法检测细胞存活率和TUNEL法流式细胞仪检测细胞凋亡；Western blot法检测肺癌细胞株中Bcl-2和p53的蛋白表达水平。结果：Real-time PCR法检测结果发现，实验组细胞中miR-214表达量明显低于对照组BEAs-2B，差异有统计学意义（P<0.05）；CCK-8结果显示，上调miR-214表达后肺癌细胞的存活率显著降低，差异有统计学意义（P<0.05）；MTT法结果显示，上调miR-214表达后肺癌细胞对抗肿瘤药物的敏感性显著降低（P<0.05）；Western blot结果表明，上调miR-214基因表达后肺癌细胞Bcl-2和p53蛋白的异常表达，与对照组相比差异有统计学意义（P<0.05）；结论：上调miR-214表达能够显著降低肺癌细胞的存活率并加速凋亡的发生；同时miR-214过表达可以调控凋亡蛋白并降低肺癌细胞对药物的敏感性。

Objective:To investigate whether microRNA-214 overexpression could affect multidrug resistance of lung cancer cells by regulating the process of apoptosis. Methods:Human small cell lung cancer cell lines and human non-small cell lung cancer cell lines were selected for H69 cell group, H446 cell group, A549 cell group and H1299 cell group; Lung epithelial cells (BEAs-2B) were served as control group. Real-time PCR was used to detect the expression of miR-214. After up-regulated microRNA-214 expression, MTT colorimetry was used to detect the sensitivity of lung cancer cells; CCK-8 assay was used to detect cell viability; TUNEL assay was used to detect the levels of apoptosis by flow cytometry; the protein expression of Bcl-2 and p53 were detected by Western blot. Results:Real-time PCR showed that the expression of miR-214 was significantly decreased in the lung cancer cell groups(P<0.05); CCK-8 results showed that miR-214 overexpression was significantly down-regulated the cell viability (P<0.05); miR-214 overexpression was significantly decreased the drug sensitivity in lung cancer cells by MTT assay(P<0.05); compared with the control group, the protein expression of Bcl-2 and p53 in lung cancer cells were significantly abnormally expressed (P<0.05). Conclusion:Up-regulation of miR-214 could decrease cell viability of the lung cancer and increase cell apoptosis, which may also regulate the apoptosis-related proteins and decrease the drug sensitivity in lung cancer cells.

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