稳定表达PLCγ1 siRNA慢病毒的构建及对人大肠癌细胞凋亡的影响  

Construction of lentivirus producing PLC-γ1 siRNA and its effect on apoptosis of human colorectal carcinomas cell lines

作　　者：谭丽[1] 罗深秋[1] 林骏[1] 

机构地区：[1]南方医科大学细胞生物教研室,广东广州510515

出　　处：《南方医科大学学报》2007年第3期255-258,共4页Journal of Southern Medical University

基　　金：广东省自然科学基金重点项目(05102580)~~

摘　　要：目的制备在人大肠癌系LoVo中稳定抑制磷脂酶C(PLC)γ1的重组慢病毒,建立PLCγ1表达降低的细胞系,以便客观研究该基因的作用。方法制备产生PLCγ1siRNA分子的重组慢病毒,在慢病毒感染LoVo细胞后以杀稻瘟菌素筛选稳定表达细胞系。应用Western-blot和RT-PCR对细胞中PLCγ1的蛋白和mRNA表达水平进行分析。应用流式细胞仪分析PLCγ1siRNA对细胞凋亡的影响。结果和结论PLCγ1siRNA显著下调了LoVo细胞中PLCγ1的表达,所构建的重组慢病毒导入的siRNA有较高的沉默效率,显著增加了5-FU诱导的凋亡。Objective To construct recombinant lentivirus that stably suppresses phosholipase C （PLC） γ1 expression in human colorectal carcinoma LoVo cells to obtain LoVo cell lines deficient in PLCγ1 for investigation of the role of PLCγ1 gene. Methods Recombinant lentivirus producing PLCγ1 siRNA were constructed to infect LoVo cells, and the stably transduced cells were selected with blasticidin. The protein and mRNA expression of PLCγ1 was examined by Western blotting and RT-PCR and the effect of the lentivirus on cell apoptosis was analyzed by flow cytometry. Results and Conclusion PLCγ1 siRNA significantly suppressed PLCγ1 expression in LoVo cells, suggesting high efficiency of gene silencing induced by the siRNA produced by the recombinant lentivirus. Concomitantly, cell apoptosis induced by 5-FU was significantly increased.

关 键 词：磷脂酶Cγ1 慢病毒 LOVO细胞 小干扰RNA 凋亡 

分 类 号：R735.34[医药卫生—肿瘤]