显性失活缺氧诱导因子-1通过下调VEGF表达促进宫颈癌细胞凋亡

Dominant-negative Variant of Hypoxia Inducible Factor 1α Down-regulates VEGF Expression and Promotes Apoptosis in SiHa Human Cervix Cancer Cell

将表达野生型缺氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)的重组质粒pcDNA3.1-full length HIF-1α,表达抑制型HIF-1α的重组质粒pcDNA3.1-dominant negative HIF-1α和空质粒pcDNA3.1稳定转染人宫颈癌SiHa细胞,研究HIF-1α对人宫颈癌SiHa细胞生物学行为的影响.采用免疫细胞化学法和Western印迹检测HIF-1α与VEGF蛋白的表达;CoCl2化学缺氧法处理细胞,采用原位缺口末端标记(TUNEL)法检测细胞凋亡情况.结果显示,显性失活HIF-1α能下调VEGF蛋白的表达,促进细胞缺氧条件下的凋亡,这提示HIF-1α可能在宫颈癌的发生发展中起作用,利用显性失活HIF-1α转染抑制HIF-1α可望成为宫颈癌治疗基因治疗的又一新途径.

The aim of the study was to evaluate the effect of hypoxia inducible factor 1α （HIF-1α） in human uterine cervix cancer cells and investigate the related mechanisms. SiHa human cervix cancer cells were stably transfected with vectors containing either a full length HIF- 1α or a dom/nant negative HIF- 1 α, and compared with the pcDNA 3.1 transfection control samples. The expressions of HIF-1α and VEGF were detected by immunocytochemistry and western blotting. The apoptosis rates of SiHa cells under CoCl2-induced chemical hypoxia were measured by TUNEL assays. Our data have shown that the dominant-negative HIF-1α inhibited the expression of VEGF and accelerates hypoxia-induced apoptosis in SiHa cells. Our results suggested that HIF-1α played an important role in the survival from hypoxia progression for cervix cancer cells, and the dominant-negative HIF-1α might be a potential reagent for the treatment of uterine cervix cancers.