中文摘要：

目的：探讨人HIF-1α短发夹（sh）RNA表达质粒对卵巢癌SKOV3细胞HIF-1α基因的表达及对紫杉醇敏感性的影响.方法：构建HIF-1α（sh）RNA表达质粒,脂质体法转染,RT-PCR和Western blot检测癌细胞HIF-1α mRNA和蛋白表达水平,MTT法检测癌细胞对紫杉醇的敏感性.结果：转染质粒48 h后,H质粒转染组SKOV3细胞紫杉醇作用24 h的IC50值明显降低（P〈0.05）,同时P-糖蛋白表达水平显著降低,差异有统计学意义（F=7.24,P=0.01）.结论：针对HIF-1α的RNAi表达质粒可下调低氧培养的卵巢癌细胞SKOV3 HIF-1α基因mRNA和蛋白及P-gp的表达,从而提高癌细胞对紫杉醇的敏感性.

英文摘要：

Objective： To investigate the effect of H/F-1α short-hairpin RNA （shRNA） expressing plasmid on ovarian cancer in vitro. Methods： Oligos for hairpin RNA expression which targeted HIF-1α gene were designed and selected based on the well-known principle, using online softwares. Annealed oligos were inserted into the pGPU6/GFP/Neo plasmid to construct recombinant plasmids. The plasmids were transfected into SKOV3 cells using lipofectmine. The expression of HIF-1α mRNA and protein was assayed by RT-PCR and Western blot respectively. Chemotherapy drug sensitivity was determined by methylthiazolyltetrazolium （ MTT ） assay. Results： At 48 hours after plasmid transfection, compared with that of Lipo group and NC group, IC50 was significantly lower in SKOV3 cells of the H plasmid transfection group undergoing a 24-hour Paclitaxel medication （ P 〈 0.05 ）. The expression of P-gp was obviously lower in SKOV3 cells of the H plasmid group than in the Lipo and NC group, with significant difference （ F = 7.24, P = 0.01 ）. Conclusion： RNAi expressing plasmid directly against HIF-1α can effectively down-regulate the expression of HIF-1α mRNA and protein in SKOV3 cells. After selectively silencing HIF-1α, hypoxia-induced expression of its target genes such as P-glycoprotein can be markedly attenuated. HIF-1α down-regulation could increase the drug sensitivity of SKOV3 cells to Paclitaxel.