抑制miR-21对非小细胞肺癌顺铂耐药细胞株A549/DDP增殖和凋亡的影响及可能机制探讨

Effects of miR-21 inhibition on A549 / DDP cisplatin resistant cell proliferation and apoptosis in non-small cell lung cancer

目的:探讨抑制miR-21表达对非小细胞肺癌耐药细胞株A549/DDP增殖和凋亡的影响及可能的机制。方法:应用Real-time PCR技术分别检测人非小细胞肺癌细胞株A549及其耐药株A549/DDP中miR-21的表达;将miR-21抑制剂(miR-21inhibitor)ASO-21转染至A549/DDP中,下调其miR-21的表达,在不同时间点使用MTT检测细胞的增殖,流式细胞仪AnnexinFITC检测细胞凋亡;使用生物信息学和miR的预测软件预测miR-21在非小细胞肺癌中的可能靶点。结果:①miR-21在A549/DDP细胞株的表达均为A549的3倍;②A549/DDP细胞株中转染ASO-21孵育,加入顺铂后,与对照组相比,其凋亡增加,增殖能力明显降低;③生物信息学软件分析后发现miR-21在非小细胞肺癌A549有多个调节靶点,一些蛋白分子可以调节miR-21的表达,组成肿瘤细胞的调节网络而影响肺癌细胞对顺铂的敏感性。结论:抑制miR-21可以促进顺铂耐药细胞A549/DDP的凋亡增加及抑制其增殖,miR-21可能成为逆转肺癌对顺铂耐药的新靶点。

Objective：To investigate the effects of miR-21 inhibition on proliferation and apoptosis of A549/DDP cells. Methods：MiR-21 expression was examined in A549 and A549/DDP cell lines. The inhibitor of miR-21（ASO-21） was transfected into A549/DDP. MTT and flow cytometry were performed to analyze the proliferation and apoptosis after transfection,respectively. Computational software and miR predicting database were used to predict the downstream effectors of miR-21 in the lung cancer cell line. Results：①miR-21 expression was approximately 3 times higher in A549/DDP compared with A549 cells. ②Apoptosis was induced and proliferation was suppressed in A549/DDP cells when miR-21 was inhibited. ③Computational softwares showed that multiple genes are possible downstream targets of miR-21. Some proteins could possibly regulate miR-21. Those genes form a regulating network via miR-21,resulting in regulating cisplatin chemoresistance. Conclusion：MiR-21 inhibition leads to apoptosis and proliferation suppression of A549/DDP cells, and it may be a therapeutic target to reverse cisplatin resistance in lung cancer treatment.