miR-409-3p靶向调控RRM2抑制鼻咽癌细胞增殖、迁移及侵袭

目的:探究微小RNA-409-3p(miR-409-3p)对鼻咽癌细胞增殖、迁移及侵袭的影响及其作用机制。方法:采用实时荧光定量聚合酶链反应(qRT-PCR)与蛋白免疫印迹法(Western blot)分别检测人永生化鼻咽上皮细胞(NP69)、鼻咽癌细胞5-8F、6-10B中miR-409-3p、核糖核苷酸还原酶M2(RRM2)的表达差异;细胞计数试剂盒(CCK-8)检测miR-409-3p过表达或RRM2过表达对6-10B细胞增殖的影响;Transwell小室实验检测miR-409-3p过表达或RRM2过表达对6-10B细胞迁移及侵袭的影响。双荧光素酶报告实验与Western blot测定miR-409-3p与RRM2在6-10B细胞中的调控关系;Western blot检测c-Myc、细胞周期蛋白1(CyclinD1)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)蛋白表达量。结果:与NP69细胞相比,5-8F、6-10B细胞中miR-409-3p的表达水平显著降低(P<0.05),RRM2的表达水平显著升高(P<0.05);miR-409-3p过表达后细胞存活率显著降低(P<0.05),迁移细胞数与侵袭细胞数显著减少(P<0.05),c-Myc、CyclinD1、MMP-2、MMP-9蛋白表达水平显著降低(P<0.05);双荧光素酶报告实验证实miR-409-3p能够靶向结合RRM2,增强miR-409-3p的表达后6-10B细胞中RRM2的表达下调(P<0.05);RRM2过表达能够明显减弱miR-409-3p过表达对6-10B细胞增殖、迁移及侵袭能力的抑制作用(P<0.05)。结论:miR-409-3p可能通过负向调控RRM2的表达从而抑制鼻咽癌细胞增殖、迁移及侵袭能力。

miR-409-3p inhibits proliferation,migration and invasion of nasopharyngeal carcinoma cells by targeted regulation of RRM2

Objective:To investigate the effect of microRNA-409-3p(miR-409-3p)on proliferation,migration and invasion of nasopharyngeal carcinoma cells and its mechanism.Methods:The expressions of miR-409-3p and RRM2 in NP69,5-8F and 6-10B cells were detected by qRT-PCR and Western blot,respectively.The cell counting kit(CCK-8)detects the effect of miR-409-3p overexpression or RRM2 overexpression on 6-10B cell proliferation.The Transwell chamber assay detects the effects of miR-409-3p overexpression or RRM2 overexpression on 6-10B cell migration and invasion.Dual luciferase reporter assay and Western blot assay for the regulation of miR-409-3p and RRM2 in 6-10B cells.Western blot was used to detect the expression of c-Myc,CyclinD1,MMP-2 and MMP-9 proteins.Results:Compared with NP69 cells and 5-8F cells,the expression level of miR-409-3p was significantly decreased in 6-10B cells(P<0.05),and the expression level of RRM2 was significantly increased(P<0.05).The cell survival rate of miR-409-3p was significantly decreased(P<0.05),the number of migrated cells and the number of invasive cells were significantly decreased(P<0.05),and the expression levels of c-Myc,CyclinD1,MMP-2 and MMP-9 were significantly decreased(P<0.05).The dual luciferase reporter assay confirmed that miR-409-3p can bind to RRM2,and the expression of RRM2 in 6-10B cells was down-regulated after enhancing the expression of miR-409-3p(P<0.05).Overexpression of RRM2 significantly attenuated the inhibitory effect of miR-409-3p overexpression on the proliferation,migration and invasion of 6-10B cells(P<0.05).Conclusion:miR-409-3p may inhibit the proliferation,migration and invasion of nasopharyngeal carcinoma cells by negatively regulating the expression of RRM2.