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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 recombinant plasmid pcDNA3.1-full length HIF-1α expressing the wild-type hypoxia inducible factor-1α (HIF-1α) and the recombinant plasmid pcDNA3.1-dominant negative HIF expressing the inhibitory HIF-1α -1αand plasmid pcDNA3.1 were transfected into SiHa cells to investigate the effect of HIF-1α on the biological behavior of human SiHa cells.Immunocytochemistry and Western blotting were used to detect the expression of HIF-1αand VEGF protein The cell apoptosis was detected by CoCl2 chemical hypoxia, and the apoptosis was detected by TUNEL method.The results showed that the dominant-negative HIF-1αmight downregulate the expression of VEGF protein and promote the hypoxia-induced apoptosis This suggests that HIF-1α may play a role in the development of cervical cancer. Inhibition of HIF-1α by dominant-negative HIF-1α transfection is expected to be another new approach for gene therapy of cervical cancer.