HPV-16 E6 siRNA与hIL-24基因联合诱导人宫颈癌Ca Ski细胞的凋亡  

HPV-16 E6 siRNA associated with hIL-24 gene induces apoptosis of human cervical cancer Ca Ski cells

作　　者：石华[1] 杨俊霞[1] 袁成福[1] 易发平[1] 魏丽丽[1] 刘革力[1] 马永平[1] 宋方洲[1] 

机构地区：[1]重庆医科大学生物化学与分子生物学教研室临床检验诊断学教育部重点实验室,重庆400016

出　　处：《肿瘤》2007年第5期348-351,共4页Tumor

基　　金：国家自然科学基金资助项目(编号:30371479);教育部春晖计划资助项目(编号:Z2004-1-55103);重庆市教委资助项目(编号:渝教科[2005]12号)

摘　　要：目的:观察HPV-16 E6 siRNA与hIL-24基因体外共转染,联合诱导人宫颈癌Ca Ski细胞凋亡的效应。方法:HPV-16 E6 siRNA与hIL-24基因的质粒载体分别以单独或联合的方式转染人宫颈癌Ca Ski细胞,随后利用RT-PCR技术检测细胞中HPV-16 E6癌基因的mRNA水平变化;Western blot分析细胞中抑癌蛋白p53水平的变化;流式细胞技术分析细胞凋亡情况。结果:经HPV-16 E6 siRNA和hIL-24转染后细胞后HPV E6癌基因的mRNA水平均下降,其中联合转染组显著下降(P<0.05);抑癌蛋白p53水平均增高,其中联合转染组显著增高,细胞凋亡率均升高,其中联合转染组显著升高(P<0.05)。结论:HPV-16 E6 siRNA与hIL-24基因分别转染宫颈癌Ca Ski细胞后,均能抑制Ca Ski细胞中HPV-16 E6癌基因的表达,使抑癌蛋白p53恢复活性,诱导宫颈癌Ca Ski细胞凋亡;两者联合则具有协同效应,能显著提高肿瘤细胞凋亡率。Objective： To evaluate the apoptosis-inducing effect of HPV-16 E6 siRNA associated with hIL-24 gene on human cervical cancer Ca Ski cells. Methods： HPV-16 E6 siRNA and hIL-24 gene eukaryotic expression vector alone or in combination were transfected into human cervical cancer Ca Ski cells. The expression level of HPV-16 E6 mRNA was detected by RT-PCR. The expression of tumor suppressor protein p53 was analyzed by western blot. The apoptosis of Ca Ski cells was analyzed by flow cytometry analysis. Results： The expression level of HPV-16 E6 mRNA decreased in Ca Ski cells in all the three groups after transfection with HPV-16 E6 siRNA and hIL-24 gene eukaryotic expression vector. The decrease was significant in co-transfection group （ P 〈 0.05 ）. The expression of tumor suppressor protein p53 and the apoptosis rate of Ca Ski cells increased in the three groups and the increase was significant in co-transfection group （P 〈 0.05 ）. Conclusion： Co-transfection of Ca Ski cells with HPV-16 E6 siRNA and hIL-24 gene inhibits the expression of tumor gene HPVE6 , recovers the activity of tumor suppressor protein p53, and induces the apoptosis of Ca Ski cells. HPV-16 E6 siRNA associated with hIL-24 gene significantly increases the apoptotic rate of Ca Ski cells. They have synergistic effects.

关 键 词：宫颈肿瘤 RNA 小分子干扰 凋亡 基因 HPV-16 E6 SIRNA 基因 hIL-24 CA Ski细胞 

分 类 号：R737.33[医药卫生—肿瘤]