siRNA表达载体对宫颈癌Caski细胞凋亡和增殖的影响-东南大学学报医学版

siRNA表达载体对宫颈癌Caski细胞凋亡和增殖的影响

单位：1. 南京医科大学第一附属医院妇产科, 江苏南京 210029;
2. 四川大学华西第二医院妇科, 四川成都 610041

分类号：R737.33; Q255; Q253

出版年·卷·期（页码）：2009·28·第三期（171-175）

摘要：

目的: 观察HPV16 E6 siRNA表达载体对宫颈癌Caski细胞凋亡和增殖的影响。方法: 利用脂质体将HPV16 E6特异性siRNA表达载体转染Caski细胞,通过荧光定量RT-PCR检测E6 mRNA的变化,流式细胞仪检测E6蛋白和细胞凋亡率的变化,MTT法和细胞克隆法检测细胞增殖的变化。结果: HPV16 E6 siRNA表达载体可明显抑制Caski细胞E6 mRNA和E6蛋白的表达,抑制率分别为89.5%和98.1%;表达载体可明显增加细胞凋亡率,抑制细胞的增殖和克隆的形成。结论: HPV16 E6 siRNA表达载体可以特异高效地抑制宫颈癌Caski细胞E6基因的表达,增加细胞的凋亡,抑制肿瘤细胞增殖和克隆的形成。

Objective To observe the effect of HPV16 E6 specific expression vector on cell proliferation and apoptosis in cervical carcinoma Caski cells.Methods HPV16 E6 siRNA expression vector and empty expression vector were transfected into Caski cells by liposome.The effects on E6 mRNA,E6 protein expression and cell apoptosis were examined respectively by real-time RT-PCR and FCM;Cell growth rate and cloning efficiency were examined by MTT and colony forming assay.Results HPV16 E6 siRNA expression vector significantly inhibited the expression levels of E6 mRNA and E6 protein,the inhibition rates being 89.5%and 98.1%respectively.Conclusion HPV16 E6 specific siRNA expression vector could specifically and efficiently inhibit the expression of E6 gene.It could induce apoptosis more effectively and inhibit cell proliferation in cervical carcinoma Caski cells.

参考文献：

[1] EFFNER M,WERNESS B A,HUIBREGTSE J M,et al.The E6 oncoprotein encoded by human papillomavirus types 16 and 18 promotes the degradation of p53[J].Cell,1990,63(6):1129-1136.
[2] BAK D.RNAi-RNA interference—an efficient way for silence[J].J Postepy Biochem,2003,49(3):136-146.
[3] CAPLEN N J,PARRISH S,IMANI F,et al.Specific inhibition of gene expression by small double-stranded RNAs in invertebrates and vertebrate systems[J].Proc Natl Acad Sci USA,2001,98:9746-9747.
[4] 李大可,彭芝兰,牛晓宇,等.RNAi表达载体对宫颈癌HPV16 E6基因表达的抑制作用[J].四川大学学报:医学版,2005,36(3):331-333.
[5] LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2 (-Delta Delta C(T)) method[J].Methods,2001,25(4):402-408.
[6] PADDISON P J,CAUDY A A,BERNSTEIN E,et al.Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells[J].Genes Dev,2002,16(8):948-958.
[7] DOORBAR J.Molecular biology of human papillomavirus infection and cervical cancer[J].Clin Sci(Lond),2006,110(5):525-541.
[8] NARISAWA-SAITO M,KIYONO T.Basic mechanisms of high-risk human papillomavirus-induced carcinogenesis: roles of E6 and E7 proteins[J].Cancer Sci,2007,98(10): 1505-1511.

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