HIF-1对人宫颈癌SiHa细胞增殖和凋亡的影响

目的观察缺氧诱导因子1α(HIF-1α)在缺氧条件下对宫颈癌SiHa细胞的增殖、生长和凋亡的影响,探讨HIF-1α在宫颈癌的发展过程中所起的作用。方法将表达全长HIF-1α的重组质粒pcDNA3.1-full length HIF-1α和空质粒pcDNA3.1转染入SiHa细胞中,分别命名为fLHIF-1α组和pcDNA3.1组,未转染细胞组命名为未转染组。应用Western Blotting和免疫细胞化学法对细胞内的HIF-1α和VEGF表达量进行检测。应用MTT方法检测不同浓度的CoCl2处理后细胞的增殖情况;采用原位缺口末端标记(TUNEL)法检测细胞的凋亡情况。结果免疫细胞化学和Western Blotting结果显示重组质粒pcDNA3.1-full length HIF-1α转染成功,转染细胞内HIF-1α及VEGF表达较未转染细胞和质粒pcDNA3.1转染细胞增高(P<0.05),显示在正常或缺氧条件下,fLHIF-1α组细胞的增殖能力均高于pcDNA3.1组和未转染组(P<0.05),凋亡率小于pcDNA3.1组和未转染组,差异均有统计学意义(P<0.05)。结论HIF-1α在CoCl2诱导的缺氧条件下可以抑制SiHa细胞的凋亡,减少缺氧对细胞的损伤以及促进细胞的增殖,提示HIF-1α在宫颈癌的发生发展中可能起着重要作用。

Effect of HIF-1alpha on the proliferation and apoptosis of uterine cervix cancer SiHa cells]

OBJECTIVE: To explore the effect of HIF-1alpha on the proliferation and apoptosis of SiHa cells in hypoxia, as well as the role of HIF-1alpha in the development of uterine cervix cancer. METHODS: The eukaryotic expression vector containing full length HIF-1alpha (pcDNA3. 1-full length HIF-1alpha) was transfected into SiHa cell, the mock plasmid (pcDNA3. 1) was also transfected into SiHa cell as the negative control. The expression of HIF-1alpha and VEGF were detected by Western Blotting and immunocytochemistry. Cell proliferation was detected by MTT colorimetric assay after the treatment of varied concentration of CoCl2, and cell apoptosis was detected by TUNEL. RESULTS: The level of HIF-1alpha and VEGF expression in the cells transfected with full length HIF-1alpha was higher than that in the untransfected group and pcDNA3. 1 group (P < 0.05), the survival ability of the cells in HIF-1alpha group was also higher than that of other two groups no matter in anoxia or in hypoxia (P < 0.05), the ratio of apoptosis was less than untransfected group and pcDNA3. 1 group (P < 0.05). CONCLUSION: HIF-1alpha inhibited cell apoptosis in CoCl2 induced hypoxia and stimulated cell proliferation. These results suggest that HIF-1alpha may play an important role in the development of uterine cervix cancer.