Livin基因沉默对人喉癌Hep2细胞凋亡和增殖及化疗敏感性的影响  

Effects of silencing Livin gene on apoptosis,proliferation,and chemosensitivity of human laryngeal carcinoma Hep2 cells

作　　者：何盛梅 李昌亚 孟令辉 赵厚育 HE Shengmei;LI Changya;MENG Linhui;ZHAO Houyu(Department of Otolaryngology,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学附属医院耳鼻咽喉科,贵州贵阳550004

出　　处：《贵州医科大学学报》2021年第2期172-178,共7页Journal of Guizhou Medical University

基　　金：贵州省科技计划项目[黔科合J字(2009)2325]。

摘　　要：目的探讨Livin基因沉默对人喉鳞癌Hep2细胞凋亡、增殖及化疗敏感性的影响。方法根据Livin基因合成3对siRNA(siRNA1~3),以Lipofectamine 2000为转染试剂转染人喉鳞癌Hep2细胞,分为Hep2-con组(control-siRNA)、Hep2-L组(Lipofectamine2000)、Hep2-s1组(Livin-siRNA1)、Hep2-s2组(Livin-siRNA2)、Hep2-s3组(Livin-siRNA3)及Hep2组(PBS),采用Western blot检测各组Hep2细胞中Livin蛋白表达水平;Hep2-s1~s3组仅保留干扰效果最好的Hep2-s1组进行后续研究,采用噻唑蓝(MTT)法检测4组细胞活力,流式细胞术检测细胞周期及凋亡率;将0.00、0.01、0.10、1.00及10.00 mg/L顺铂(DDP)作用于上述4组细胞后采用平板克隆形成实验、MTT法及流式细胞仪检测各组细胞对化疗敏感性的影响。结果与Hep2组比较,Hep2-s1组人喉鳞癌Hep2细胞增殖受到抑制、细胞凋亡率升高(P<0.05);与Hep2组比较,加入不同浓度DDP后Hep2-s1组Hep2细胞克隆形成率降低、集落抑制率升高、细胞存活率下降(P<0.05);与Hep2组比较,5 mg/L顺铂作用后Hep2-s1组Hep2细胞凋亡率和生长抑制率随时间延长增加,各检测点细胞凋亡率和生长抑制率升高(P<0.05)。结论 RNAi沉默Hep2细胞中Livin表达,可抑制肿瘤细胞增殖、诱导细胞凋亡及增加肿瘤细胞对顺铂化疗的敏感性。Objective To investigate the effect of silencing Livin gene on the apoptosis,proliferation and chemosensitivity of human laryngeal squamous cell carcinoma Hep2 cells. Methods Three pairs of siRNA( siRNA 1 ~ 3) against human Livin gene were synthesized. Hep2 cells were transfected with control siRNA and three siRNA against Livin gene,respectively. Lipofectamine 2000 was used as a transfection agent. Cells were divided into Hep2-con group( control-siRNA), Hep2-L group( Lipofectamine2000),Hep2-s1 group( Livin-siRNA1),Hep2-s2 group( Livin-siRNA2),Hep2-s3 group( Livin-siRNA3) and Hep2 group( PBS). Western blot was used to detect the expression level of Livin protein in Hep2 cells. In the Hep2-s1 ~ s3 group,only the Hep2-s1 group with the maximum silencing was retained for the follow-up study. The thiazole blue( MTT) assay was used to measure the cell viability. Flow cytometry was used to detect the cell cycle and apoptosis rate. Cells were treated with cisplatin( DDP) at 0. 00,0. 01,0. 10,1. 00,and 10. 00 mg/L concentration for colony formation experiment and MTT assay to detect the effects of silencing Livin gene on the sensitivity of chemotherapy in Hep G2 cells. Results Compared with the Hep2 group,Hep2-s1 group had decreased cell proliferation and increased apoptosis rate( P < 0. 05). In response to DDP treatment at indicated concentration,Hep2-s1 group exhibited reduced colony formation,increased inhibition rate in colony formation and decreased cell viability relative to Hep2 group( P < 0. 05). When compared with the Hep2 group,the apoptotic rate and growth inhibition rate in Hep2-s1 group were increased after treatment with 5 mg/L cisplatin. The apoptosis rate and growth inhibition rate were increased at each detection point( P <0. 05). Conclusion RNAi-mediated Livin silencing in Hep2 cells can inhibit cell proliferation,induce cell apoptosis,and increase the sensitivity of cells to cisplatin chemotherapy.

关 键 词：喉 顺铂 肿瘤 LIVIN基因 RNA干扰 敏感性 

分 类 号：R767[医药卫生—耳鼻咽喉科]