靶向HPV18<EM>E</EM>6基因siRNA对HeLa细胞p21、VEGF、Bax 和<EM>Bcl</EM>-2基因的影响

doi:10.3971/j.issn.1000-8578.2009.04.001

靶向HPV18E6基因siRNA对HeLa细胞p21、VEGF、Bax 和Bcl-2基因的影响

Influence of HPV18E6 Small Interfering RNA on p21,VEGF,Bax and Bcl-2 Genes in HeLa Cells

摘要

摘要: 目的探讨针对人乳头瘤病毒18型E6基因的特异性小干扰RNA（siRNA）对宫颈癌HeLa细胞p21、VEGF、Bax和Bcl-2基因的影响。方法采用脂质体法将特异性siRNA瞬时转染HeLa细胞，半定量RT-PCR检测siRNA转染后HeLa细胞中p21、血管内皮生长因子（VEGF）、Bax和Bcl-2 mRNA的变化，免疫组化检测HeLa细胞中p21和VEGF蛋白的变化。结果 RT-PCR检测结果显示转染后24、48、 72h p21和Bax mRNA的表达与转染前比较均有升高。转染后24、48、72h VEGF和Bcl-2 mRNA的表达与转染前比较均有降低。免疫组化检测结果显示转染后48、72h p21蛋白的表达与转染前比较均有升高。转染后48、72h VEGF蛋白的表达与转染前比较均有降低。结论靶向HPV18 E6基因的siRNA可有效干扰宫颈癌HeLa细胞中p21、VEGF、 Bax和Bcl-2基因的表达，从而抑制细胞增殖。

Abstract: Objective To investigate the influence of HPV18E6 small interfering RNA on p21,VEGF,Bax and Bcl-2 genes in human cervical cancer cell line HeLa. Methods The specific siRNA was transfected into HeLa cells by lipofectamine.Expressions of p21,VEGF,Bax and Bcl-2 mRNA were examined by RT-PCR.Immunochemitry was used to evaluate the expression of p21 and VEGF protein. Results Compared with control group,the expressions of p21 and Bax mRNA were increased after siRNA transfection,while the mRNA levels of VEGF and Bcl-2 were decreased significantly.Immunochemical results showed higher expression of p21 and lower expression of VEGF protein. Conclusion HPV18 E6 small interfering RNA can after the expression of p21,VEGF,Bax and Bcl-2 genes in the human cervical cancer cell line HeLa and inhibit the proliferation of cells.

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