EBV下调miR-34c促进鼻咽癌细胞增殖、迁移的作用及机制研究

Role of miR-34c downregulated by EBV in promoting proliferation and migration of nasopharyngeal carcinoma cells and its mechanism

目的:探究爱泼斯坦-巴尔病毒(EBV)下调miR-34c促进鼻咽癌细胞增殖、迁移、侵袭的作用及机制。方法:采用qPCR检测EBV阴性鼻咽癌SUNE1、CNE2、HK1细胞和EBV阳性C666-1细胞中miR-34c表达水平。构建miR-34c模拟物以分析miR-34c对鼻咽癌细胞生物学功能的影响。使用CCK-8、划痕愈合试验以及Transwell细胞侵袭试验检测鼻咽癌细胞的增殖、迁移和侵袭能力。Western blotting测定细胞中迁移、侵袭相关蛋白以及Erk1/2信号通路蛋白的表达水平。结果:miR-34c在EBV阳性C666-1细胞表达下调(P<0.05)。CCK-8结果显示,miR-34c组C666-1细胞在16、24、48 h的活力明显低于EBV组(P<0.01)。划痕试验和Transwell试验结果显示,与miR-34c组比较,EBV组C666-1细胞迁移和侵袭能力均明显增强(P<0.01)。Western blotting结果显示,与miR-34c组比较,EBV组C666-1细胞迁移和侵袭相关蛋白Vimentin、Snail、MMP-2、MMP-3及Erk1、Erk2蛋白表达均明显升高(P<0.01)。结论:miR-34c在EBV阳性鼻咽癌细中表达下调,EBV下调miR-34c可增强鼻咽癌细胞增殖、侵袭和迁移能力及激活Erk1/2信号通路。

Objective:To investigate the role of miR-34c downregulated by Epstein-Barr virus(EBV)in promoting the proliferation,migration,and invasion of nasopharyngeal carcinoma cells and its mechanism.Methods:The expression level of miR-34c in EBV-negative nasopharyngeal carcinoma SUNE1,CNE2,HK1 cells,and EBV-positive C666-1 cells was detected by qPCR.MiR-34c mimics were constructed to analyze the influence of miR-34c on the biological function of nasopharyngeal carcinoma cells.CCK-8,scratch healing assay,and Transwell cell invasion assay were applied to detect the proliferation,migration,and invasion ability of nasopharyngeal carcinoma cells.Western blotting was employed to determine the expression levels of migration and invasion-related proteins,and Erk1/2 signaling pathway proteins in cells.Results:MiR-34c was downregulated in EBV-positive C666-1 cells(P<0.05).CCK-8 results showed that the viability of C666-1 cells in the miR-34c group was significantly lower than that in the EBV group at 16,24,and 48 hours(P<0.01).The scratch assay and Transwell assay results indicated that compared with the miR-34c group,the migration and invasion ability of C666-1 cells in the EBV group were significantly enhanced(P<0.01).Western blotting results showed that compared with the miR-34c group,the expression of migration and invasion-related proteins Vimentin,Snail,MMP-2,MMP-3,and Erk1 and Erk2 proteins of C666-1 cells in the EBV group were significantly increased(P<0.01).Conclusions:MiR-34c is downregulated in EBV-positive nasopharyngeal carcinoma cells,and miR-34c downregulated by EBV can enhance the proliferation,invasion,and migration capacities of nasopharyngeal carcinoma cells and activate the Erk1/2 signaling pathway.