沉默A20基因对人喉鳞癌细胞系Hep-2增殖、周期、侵袭能力的影响  

Effects of silencing A20 on proliferation,cell cycle,and invasive of human laryngeal squamous cell carcinoma Hep-2 cells

作　　者：罗德艳 冯俊 彭涛 唐一萍 LUO Deyan;FENG Jun;PENG Tao;TANG Yiping(Nanchong Central Hospital,Nanchong 637000,China)

机构地区：[1]南充市中心医院

出　　处：《山东医药》2020年第3期13-16,共4页Shandong Medical Journal

基　　金：四川省科技厅重点研发计划项目(2018FZ0116);四川省教育厅自然科学重点项目(18ZA203);南充市科技局应用技术研究与开发基金项目(16YFZJ0021)

摘　　要：目的观察沉默肿瘤坏死因子α诱导蛋白3(A20)基因对人喉鳞癌细胞系Hep-2增殖、周期、侵袭能力的影响。方法取对数生长期Hep-2细胞,接种至6孔板,待细胞融合达80%~90%时进行分组,实验共分为A20siRNA组、NC siRNA组、对照组,A20 siRNA组、NC siRNA组应用Lipofectamine RNAi MAX转染试剂分别转染A20siRNA、NC siRNA,转染操作严格按照试剂盒说明书操作,对照组不做任何处理。转染后48 h,采用qRT-PCR法和Western blotting法分别检测A20 mRNA、蛋白,CCK8实验、平板克隆实验检测细胞增殖能力,流式细胞术检测细胞周期,Transwell小室测定细胞侵袭能力。结果与NC siRNA组及对照组比较,A20 siRNA组A20 mRNA、蛋白表达降低,24、48、72 h的OD值降低,细胞G0/G1期比例升高,细胞S期、G2/M期比例降低,穿膜细胞数降低(P均<0. 05)。结论沉默A20基因能够抑制Hep-2细胞增殖和侵袭,缩短细胞周期。Objective To observe the effects of silencing tumor necrosis factor-α-inducible protein 3( A20) gene on the proliferation,cell cycle,and invasion of human laryngeal squamous cell carcinoma cell line Hep-2. Methods Hep-2 cells in the logarithmic phase were inoculated into 6-well plate,and then they were divided into the A20 siRNA group,NC siRNA group,and control group when the cell fusion reached 80%-90%. Cells in the A20 siRNA group and NC siRNA group were transfected with A20 siRNA and NC siRNA,respectively,by Lipofectamine RNAi MAX transfection reagent.The transfection operation was strictly in accordance with the instructions of the kit,and the control group was not treated.At 48 h after transfection,the expression of A20 mRNA and protein in Hep-2 cells were detected by qRT-PCR and Western blotting,the proliferation ability of Hep-2 cells was detected by CCK8 and plate cloning experiment,the cell cycle of Hep-2 cells was detected by flow cytometry,and the invasion ability of Hep-2 cells was detected by Transwell cell chamber. Results Compared with the NC siRNA group and control group,the expression of A20 mRNA and protein decreased,the OD values decreased at 24,48 and 72 h,the proportion of cells in the G0/G1 phase increased,the proportion of cells in the S phase and G2/M phase decreased,and the number of cells penetrating the membrane decreased in the A20 siRNA group( all P < 0. 05). Conclusion Silencing A20 gene can inhibit the proliferation and invasion of Hep-2 cells and shorten the cell cycle.

关 键 词：喉鳞癌 HEP-2细胞 A20基因 细胞增殖 细胞周期 细胞侵袭 

分 类 号：R739.65[医药卫生—肿瘤]