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

作　　者：赵凤艳[1] 唐彬秩[2] 欧阳运薇 母得志[1] 毛萌[1] 张林[1] 庞秋霞[3] 屈艺[1] 

机构地区：[1]四川大学华西第二医院儿科,成都610041 [2]四川大学华西基础医学与法医学院生物化学与分子生物学研究室,成都610041 [3]陕西省延安大学生物化学教研室,延安716000

出　　处：《中国生物化学与分子生物学报》2008年第3期237-243,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金(No.30570623);国家教育部博士点基金(No.20050610094)资助~~

摘　　要：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.

关 键 词：缺氧诱导因子-1 血管内皮生长因子 SIHA细胞 凋亡 

分 类 号：R739[医药卫生—肿瘤] R737.25[医药卫生—临床医学]