Bmi-1基因RNAi对喉癌细胞Hep-2生物学行为的影响

目的:构建Bmi-1基因的RNAi表达载体,观察其对喉癌细胞Hep-2增殖、侵袭能力的影响。方法:利用慢病毒表达体系pHelper1.0/pHelper2.0/pGCL2GFP,构建Bmi-1基因的RNAi重组质粒vshRNA-Bmi-1。实时定量PCR和蛋白质印迹法分别检测稳定转染vshRNA-Bmi-1后喉癌细胞中Bmi-1mRNA及蛋白的表达;克隆形成实验及小室侵袭实验检测喉癌细胞增殖和侵袭能力的变化。结果:稳定转染vshRNA-Bmi-1后Hep-2细胞Bmi-1mRNA表达均明显下降,Hep-2细胞中Bmi-1蛋白表达明显下降。抑制率分别为79%及88%。Bmi-1干扰后Hep-2细胞增殖减慢、侵袭能力减弱。结论:成功构建Bmi-1基因的RNAi表达载体,vshRNA-Bmi-1重组质粒明显下调Bmi-1mRNA和蛋白在喉癌细胞Hep-2中的表达。Bmi-1基因的RNAi能抑制喉癌细胞Hep-2的增殖和侵袭能力。

Effects of Bmi-1 RNAi gene on laryngeal carcinoma Hep-2 cells

Objective:To construct the Bmi-1 RNAi expression vector and investigate its influence on the proliferation and invasiveness of laryngeal carcinoma Hep-2 cells.Method:The recombinant vshRNA-Bmi-1 plasmid of Bmi-1 RNAi was constructed by the lentiviral expression system,pHelper1.0/pHelper2.0/pGCL2GFP.Bmi-1mRNA and protein expressions of stably transfected laryngeal carcinoma cells were identified respectively by real-time PCR and Western blot analyses.The changes of the proliferation and invasiveness of laryngeal carcinoma were detected by clone formation test and an invasion assay.Result:The Bmi-1 mRNA expressions of stably transfected laryngeal carcinoma Hep-2 cells were significantly decreased.The expression of Bmi-1 protein in laryngeal carcinoma Hep-2 cells was significantly decreased.And the inhibitory rates were 79% and 88% respectively.Whereas the proliferation and invasiveness of Hep-2 cells were significantly reduced.Conclusion:The Bmi-1 RNAi expression vector were constructed successful.Reduced Bmi-1 expression of Hep-2 cells demonstrated the role of Bmi-1 RNAi in restraining proliferation and invasiveness of laryngeal carcinoma cells.