敲除OIP5-AS1基因通过miR-7-5p/PARP1轴增强非小细胞肺癌细胞的化学敏感性

Knockout of long-chain non-coding RNA OIP5-AS1 enhances chemosensitivity of non-small cell lung cancer cells via miR-7-5p/PARP1 Axis

目的探讨OIP5-AS1基因对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞化学敏感性的作用及分子机制。方法采用CCK-8法检测正常肺细胞系BEAS-2B、肺癌细胞系A549和顺铂耐药肺癌细胞A549/DDP的半抑制浓度(IC50)值。应用RT-qPCR检测OIP5-AS1、miR-7-5p和PARP1的表达水平。生物信息学检测OIP5-AS1、miR-7-5p和PARP1的靶向关系,用荧光素实验进一步验证。Western blot法检测相关蛋白的表达;流式细胞术检测细胞凋亡情况;克隆形成检测细胞生长情况。结果A549/DDP细胞中IC50值最高,且OIP5-AS1高表达,miR-7-5p低表达。miR-7-5p是OIP5-AS1的靶向调节基因之一,且为负调控关系。敲除OIP5-AS1抑制A549/DDP细胞的生长并增强化学敏感性,促进A549/DDP细胞凋亡。敲除OIP5-AS1下调PARP1可以增强A549/DDP细胞的化学敏感性,抑制NSCLC的生长和凋亡。结论OIP5-AS1通过上调miR-7-5p或下调PARP1,增强A549/DDP细胞的化学敏感性,抑制NSCLC的生长和凋亡。

Purpose To investigate the effect of OIP5-AS1 on the chemosensitivity of non-small cell lung cancer(NSCLC)cells and its molecular mechanism.Methods The CCK-8 method was used to detect the half maximal inhibitory concentration(IC50)value of normal lung cell line BEAS-2B,lung cancer cell line A549 and cisplatin-resistant cell A549/DDP.The expression levels of OIP5-AS1,miR-7-5p and PARP1 were detected by RT-qPCR.The bioinformatics detection of the targeting relationship of OIP5-AS1,miR-7-5p and PARP1 was further verified by fluorescein experiments.Western blot was applied to detect the expression of related proteins.Flow cytometry was used to detect cell apoptosis.Colony formation detected cell growth.Results A549/DDP cells had the highest IC50 values,and OIP5-AS1 was highly expressed and miR-7-5p was lowly expressed.miR-7-5p was one of the targeting regulatory genes of OIP5-AS1,and had a negative regulatory relationship.Knockout of OIP5-AS1 inhibited the growth of A549/DDP cells and enhanced chemosensitivity and promoted apoptosis of A549/DDP cells.Knockdown of OIP5-AS1 down-regulation of PARP1 enhanced the chemosensitivity of A549/DDP cells and inhibits the growth and apoptosis of NSCLC.Conclusion OIP5-AS1 enhances the chemosensitivity of A549/DDP cells and inhibits non-small cell lung cancer by up-regulating miR-7-5p or down-regulating PARP1.