miR-148a靶向HMGB3抑制非小细胞肺癌细胞迁移、侵袭和增强顺铂抗肿瘤效应的机制研究

Mechanism of miR-148a on migration and invasion of non-small cell lung cancer cells and enhancing effection of DDP anti-tumor by targeting HMGB3

目的:探讨miR-148a靶向HMGB3抑制非小细胞肺癌细胞迁移和侵袭的作用机制。方法:采用RT-qPCR和Western blot检测非小细胞肺癌组织和癌旁组织标本中miR-148a和HMGB3相对表达水平;以A549、H1299细胞为研究对象,Transwell和MTT法分别检测过表达miR-148a、敲低HMGB3和DDP干预对细胞迁移、侵袭及细胞活力的影响;TargetScan在线预测、双荧光素酶报告基因实验和Western blot验证miR-148a与HMGB3的靶向关系;miR-148a过表达或敲减HMGB3并联合DDP,Transwell和MTT法检测细胞迁移、侵袭和细胞活力。结果:非小细胞癌组织中miRNA-148a表达明显下调(P<0.05),HMGB3在mRNA和蛋白水平表达均明显上调(P<0.05),miR-148a与HMGB3表达呈负相关(r=-0.8568,P<0.0001);过表达miR-148a或敲低HMGB3能明显抑制细胞迁移和侵袭(P<0.05),过表达miR-148a或敲低HMGB3联合DDP干预,细胞活力明显下降(P<0.05);TargetScan在线预测、双荧光素酶报告基因实验和Western blot检测表明miR-148a能调控HMGB3表达。结论:miR-148a在非小细胞肺癌组织中表达下调,HMGB3是miR-148a的靶基因,过表达miR-148a能抑制HMGB3表达从而抑制非小细胞肺癌细胞A549、H1299迁移和侵袭并增强顺铂抗肿瘤效应。

Objective:To investigate the effect of miR-148a on migration and invasion of non-small cell lung cancer cells by directly targeting HMGB3.Methods:The expression of miR-148a and HMGB3 in non-small cell lung cancer and para-cancerous tissues was detected by RT-qPCR and Western blot.Cell migration,invasion and cell viability of A549 and H1299 cells after over-expression of miR-148a,knockdown of HMGB3 or DDP intervention were detected by Transwell and MTT assay,respectively.The regulating relationship between miR-148a and HMGB3 was verified by TargetScan online prediction,dual luciferase reporter system and Western blot.Cell migration,invasion and cell viability of A549 and H1299 cells after over-expression of miR-148a and HMGB3 combined with DDP intervention were detected by Transwell and MTT assay,respectively.Results:The expression level of miR-148a in non-small cell lung cancer tissues was down-regulated ( P <0.05),while HMGB3 had the opposite trend in the tissues ( P <0.05).The expression of miR-148a was negatively correlated with HMGB3 ( r =-0.856 8,P <0.000 1).The migration and invasion of A549 and H1299 cells were inhibited after over-expression of miR-148a or knockdown of HMGB3.Cell viability decreased significantly after over-expression of miR-148a or knockdown of HMGB3 combined with DDP ( P <0.05).The result of TargetScan online prediction,dual luciferase assay and Western blot showed that miR-148a negatively regulated the expression of HMGB3.Conclusion:miR-148a is down-regulated in non-small cell lung cancer tissues,and over-expression of miR-148a can inhibit the migration and invasion and enhance the effect of DDP antitumor of non-small cell lung cancer cells A549 and H1299 by directly targeting HMGB3.