miR-142-5p对非小细胞肺癌A549顺铂敏感性的影响及机制

The effect and mechanism of miR-142-5p on cisplatin sensitivity in non-small cell lung cancer A549

目的探讨miR-187对胶质瘤U251细胞增殖、侵袭、迁移和凋亡的影响及其机制。方法将体外培养的U251细胞分为Con组(未处理)、NC组(转染miR-NC)、miR-187组(转染miR-187 mimics)。采用实时定量PCR检测miR-187的表达,MTT法、Transwell小室实验和流式细胞仪分别检测细胞增殖、迁移、侵袭和凋亡,实时定量PCR和Western blot检测S100A4 mRNA和蛋白的表达,双荧光素酶报告基因实验检测miR-187和S100A4的靶向关系。结果上调miR-142-5p表达可以显著阻滞A549细胞周期、抑制细胞增殖,促进细胞凋亡,激活Akt/mTOR通路活性,抑制LC3B、Beclin-1的表达,造成细胞自噬水平下降。结论 miR-142-5p协同顺铂促进A549细胞凋亡,与细胞自噬有关,miR-142-5p有望成为非小细胞肺癌临床治疗的潜在作用靶点。

Objective To study the regulatory effect of miR-142-5 p on cisplatin chemosensitivity in non-small cell lung cancer cells and to provide theoretical basis for clinical treatment.Methods The expression level of miR-142-5 p was up-regulated or down-regulated by transfection technique,the cell proliferation,cycle and apoptosis were detected by CCK-8 and flow cytometry.Real-time PCR was used to detect miR-142-5 p expression.Western blot was used to detect the expressions of Akt/mTOR pathway related proteins,autophagy markers LC3 BⅡ/Ⅰand Beclin-1.Immunofluorescence was used to observe the punctiform aggregation of LC3 B protein.Results Up-regulation of miR-142-5 p expression level inhibited significantly the cell cycle and proliferation of A549 cells,promoted the cell apoptosis,activated Akt/mTOR pathway and inhibited the expression of LC3 B and Beclin-1,resulting in declined level of cell autophagy.Conclusion miR-142-5 p is expected to be a potential target for clinical treatment of non-small cell lung cancer.