lncRNA DANCR通过调控miR-214-5p在宫颈癌细胞增殖及侵袭中的作用研究

Role of lncRNA DANCR in the proliferation and invasion of cervical cancer cells by regulating miR-214-5p

目的探讨长链非编码RNA(lncRNA)分化拮抗非蛋白编码RNA(DANCR)通过调控miR-214-5p对宫颈癌细胞增殖和侵袭的影响及作用机制。方法用实时荧光定量逆转录聚合酶链反应(qRT-PCR)分别检测DANCR在宫颈癌组织和细胞系中的表达情况,向HeLa细胞转染过表达DANCR质粒(过表达DANCR组),并设置Vector组作为对照,检测DANCR的相对表达情况,进一步向过表达DANCR的HeLa细胞中转染miR-214-5p模拟物(DANCR+miR-214-5p组)和miR阴性对照物(DANCR+miR NC组),并设置Vector组作为空白对照组,检测miR-214-5p在各组中的相对表达情况;用细胞计数(CCK-8)法和Transwell实验检测DANCR对宫颈癌细胞增殖和侵袭的影响;用双荧光素酶报告基因实验检测DANCR与miR-214-5p的结合关系。结果DANCR在癌旁组织和宫颈癌组织中的相对表达分别为1.00±0.09,1.79±0.15,差异有统计学意义(P<0.01)。与人宫颈上皮永生化细胞H8相比,DANCR在人宫颈癌细胞系SiHa、HeLa、MS751和HT-3中表达均显著上调(均P<0.01);与Vector组相比,过表达DANCR促进了HeLa细胞的增殖(均P<0.05)。对照组和过表达DANCR组的侵袭细胞数分别为15.33±1.51和28.33±2.76,过表达DANCR组的细胞侵袭能力显著高于对照组(P<0.01)。Vector组和过表达DANCR组中miR-214-5p的相对表达水平分别为1.00±0.09,0.38±0.03,差异有统计学意义(P<0.01)。Vector组、DANCR+miR NC组和DANCR+miR-214-5p组miR-214-5p相对表达分别为1.00±0.10,0.37±0.03,0.84±0.08,组间比较差异均有统计学意义(均P<0.01)。对照组和DANCR+miR NC组以及DANCR+miR-214-5p组的细胞侵袭数分别为13.66±1.33,19.66±1.94,14.33±1.42,与对照组相比,DANCR+miR NC组的细胞侵袭能力显著增强(P<0.05),与DANCR+miR NC组相比,DANCR+miR-214-5p组的细胞侵袭能力显著降低(P<0.05)。结论DANCR通过调控miR-214-5p促进宫颈癌细胞的增殖及侵袭。

Objective To investigate the effects and mechanisms of long-chain non-coding RNA(lncRNA)differentiation antagonizing non-protein coding RNA(DANCR)on the proliferation and invasion of cervical cancer cells by regulating miR-214-5 p.Methods Real-time fluorescent quantitative(qRT-PCR)was used to detect the expression of DANCR in cervical cancer tissues and cell lines,and the overexpressed DANCR plasmid(overexpressing DANCR group)was transfected into HeLa cells.The Vector group was used as the control to detect the relative expression of DANCR.Further,miR-214-5 p mimic was transfected(DANCR+miR-214-5 p group)and miR negative control(DANCR+miR NC group)was transfected into HeLa cells overexpressing DANCR,and Vector group was set up as blank control group to detect the relative expression of miR-214-5 p in each group.The effects of DANCR on the proliferation and invasion of cervical cancer cells were detected by cell counting kit(CCK-8)and Transwell experiments.The binding relationship between DANCR and miR-214-5 p was detected by dual-luciferase reporter gene experiment.Results The relative expression levels of DANCR in paracancerous tissues and cervical cancer tissues were 1.00±0.09,1.79±0.15,with significant difference(P<0.01).Compared with the human cervical epithelial immortalization cell line H8,the expression levels of DANCR in the human cervical cancer cell lines Si Ha,HeLa,MS751 and HT-3 were significantly increased(all P<0.01).Compared with Vector group,the overexpression of DANCR promoted the proliferation of HeLa cells(P<0.05).The number of invasive cells in control group and overexpressed DANCR group were 15.33±1.51 and 28.33±2.76,the cell invasion ability of the overexpressed DANCR group was significantly higher than that of control group(P<0.01).The relative expression levels of miR-214-5 p in Vector group and overexpression of DANCR group were 1.00±0.09,0.38±0.03,with significant difference(P<0.01).The relative expression levels of miR-214-5 p in Vector group,DANCR+miR NC group,and DANCR+miR-214-5 p group were 1.00±0.10,0.37±0.03,0.84±0.08,with significant difference(P<0.01).The cell invasion numbers in control group,DANCR+miR NC group,and DANCR+miR-214-5 p group were 13.66±1.33,19.66±1.94 and 14.33±1.42,compared with the control group,the cell invasion ability of the DANCR+miR NC group was significantly increased(P<0.05).Compared with DANCR+miR NC group,the cell invasion ability of the DANCR+miR-214-5 p group was significantly decreased(P<0.05).Conclusion DANCR promotes the proliferation and invasion of cervical cancer cells by regulating miR-214-5 p.