摘要
目的:研究真核起始因子4E(eukaryotic initiation factor 4E,eIF4E)-小干扰RNA(small interfering RNA,siRNA)对舌鳞状细胞癌(简称鳞癌)细胞粘附和运动能力的影响。方法:舌鳞癌细胞SCC25感染靶向eIF4E干扰的慢病毒,用实时定量聚合酶链反应(quantitative real time polymerase chain reaction,qRT-PCR)和Western blot方法检测沉默效果,噻唑蓝(methylthiazolyldiphenyl-tetrazolium bromide,MTT)方法测定细胞增殖和粘附能力,划痕愈合实验测定细胞运动能力,Transwell实验测定细胞侵袭及迁移能力,Western blot检测细胞中间质标志物神经性钙粘蛋白(N-cadherin)、上皮标志物上皮钙粘蛋白(E-cadherin)和基质金属蛋白酶(matrix metalloprotease,MMP)-9、MMP-2蛋白表达情况。结果:靶向eIF4E干扰的慢病毒感染后的SCC25细胞中eIF4EmRNA和蛋白表达水平均降低。沉默eIF4E后的SCC25细胞的增殖活性降低,细胞粘附能力降低,细胞运动能力降低,侵袭和迁移能力降低,上皮标志物E-cadherin蛋白表达水平升高,间质标志物N-cadherin蛋白表达水平降低,细胞中MMP-9、MMP-2蛋白表达水平降低。结论:eIF4E-siRNA能够下调舌鳞癌细胞中eIF4E的表达,抑制舌鳞癌细胞粘附、运动、侵袭、迁移能力。
Objective:To study the effect of eIF4 EsiRNA on adhesion and movement ability of tongue squamous cell carcinoma cells.Methods:Tongue squamous cell carcinoma cell line SCC25 was infected targeting eIF4 Einterferred lentivirus.The effect of silence was detected by qRT-PCR and Western blot.Cell proliferation and adhesion were detected by MTT assay.Cell motility was detected by scratch healing test.Cell invasion and migration were measured by Transwell chamber.The expression of N-cadherin,E-cadherin,and matrix degrading enzyme MMP-9and MMP-2proteins were detected by Western blot.Results:The levels of eIF4 EmRNA and protein expression in SCC25 cells infected with eIF4 Einterfered with lentivirus were decreased.After silencing eIF4 E,the proliferation activity of SCC25 cells was decreased,cell adhesion,cell motility,and the ability to invasion and migration were decreased,the level of E-cadherin protein in the epithelial marker was elevated,the level of N-cadherin protein in the interstitial marker was reduced,and the levels of MMP-9and MMP-2protein protein in the cells were decreased.Conclusion:eIF4EsiRNA can downregulate the expression of eIF4 Ein tongue squamous cell carcinoma cells,and inhibit the adhesion,movement,invasion,and migration of tongue squamous cell carcinoma cells.
引文
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