沉默缺氧诱导因子-1α对卵巢癌化疗敏感性的影响

Inhibition of HIF-1αexpression on chemosensitivity of ovarian cancer cell line SKOV3

目的观察沉默缺氧诱导因子HIF-1α对卵巢癌细胞化疗敏感性的相关影响并探讨其机理。方法构建HIF-1α基因短发夹RNA真核表达质粒,并转染卵巢癌细胞SKOV3(人卵巢浆液性囊腺癌细胞株)。实验可分为空白组,非特异对照组(转染pNonspecific-siRNA,即SKOV3NS),目的siRNA组(转染pHIF-siRNA,即SKOV3siRNA)3组,用RT-PCR及Western Blot法分别检测各组HIF-1α基因的mRNA和蛋白的表达水平。细胞经过20μmol/L顺铂作用后,用MTT法检测细胞生长抑制率,流式细胞术(FCS)检测细胞凋亡率。结果 SKOV3siRNA转染组HIF-1α基因在mRNA水平和蛋白水平表达均明显降低,SKOV3siRNA组肿瘤细胞的生长抑制率和细胞凋亡率均明显增高(P<0.05)。结论 RNA干扰技术沉默HIF-1α能够有效地抑制卵巢癌SKOV3细胞HIF-1α基因表达,增强其对化疗药物顺铂的敏感性。

Objective Observe silence hypoxia-inducible factor（HIF-1α） on ovarian cancer cell chemosensitivity and to explore its mechanism.Methods Construction of HIF-1α shRNA eukaryotic expression vector and transfected into ovarian cancer cells SKOV3.These vectors were divided into 3 groups： SKOV3,SKOV3NS and SKOV3siRNA.They were detected by RT-PCR and Western Blot method HIF-1α gene mRNA and protein levels.Inhibition of cell growth rate was detected by MTT,cell apoptosis was detected by flow cytometry（FCS）.Results The SKOV3siRNA group in HIF-1αgene expression at the mRNA and protein levels were significantly lower.The SKOV3siRNA tumor cell growth inhibition rate and apoptosis were significantly higher（P0.05）.Conclusion RNA interference silencing HIF-1αcan effectively suppress ovarian carcinoma cell line SKOV3 HIF-1αgene expression,to enhance its sensitivity to chemotherapy drug cisplatin.