摘要
目的探讨低氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)特异性小片段干扰RNA表达载体pSU-PERH1-siHIF1α对低氧诱导因子-1α和血管内皮生长因子(vas-cular endothelial growth factor,VEGF)mRNA表达的抑制作用。方法构建HIF-1α特异性小片段干扰RNA表达载体pSUPERH1-siHIF1α,通过脂质体Lipofectamine2000分别介导pSUPERH1-siHIF1α转染低氧状态下培养的人低分化鼻咽鳞癌细胞(nasopharyngeal carcinoma cell line,CNE2),对照组转染pSUPER空载体,采用半定量RT-PCR方法检测HIF-1αmRNA和VEGFmRNA的表达。结果转染pSUPERH1-siHIF1α的CNE2细胞中HIF-1αmRNA和VEGFmRNA的表达较对照组分别降低79.4%和65.7%。结论HIF-1α特异性小片段干扰RNA表达载体pSUPERH1-siHIF1α能明显抑制HIF-1αmRNA和VEGFmRNA的表达。
Objective To evaluate the inhibition effect of the HIF-1α specific siRNA expression vector pSUPER^H1-SiHIF1α on HIF-1α and VEGF mRNA expression in vitro. Methods The HIF-1α specific siRNA expression vector pSUPER^H1-SiHIF1α was constructed by gene recombination and then transfeeted into a human CNE2 cell line euhured under hypoxia conditions （1%） with Lipofectamine 2000. The transfection of the pSUPER vector was used as a control. HIF-1α and VEGF mRNA expression were detected by semi-quantitative RT-PCR. Results The expression of HIF-1α and VEGF mRNA of CNE2 transfected with pSUPER^H1-SiHIF1α was dramatically lower than that of the control transfection. Conclusion HIF-1α specific siRNA expression plasmid pSUPER^H1-SiHIF1α can specifically down-regulate the expression of HIF-1α and VEGF mRNA in CNE2 under hypoxia conditions.
出处
《眼视光学杂志》
2007年第4期228-231,共4页
Chinese Journal of Optometry & Ophthalmology
基金
国家自然科学基金资助项目(030471853)
关键词
RNA干扰
低氧诱导因子-1
血管内皮生长因子
低分化鼻咽鳞癌细胞
RNA interference
hypoxia inducible growth factor-1
vascular endothelial growth factor
nasopharyngeal carcinma cell line
