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

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.