【摘要】 目的研究靶向干扰hif-1α基因表达后,对HeLa细胞增殖及其对顺铂敏感度的影响。方法构建针对hif-1α基因的干扰载体并转染HeLa细胞,G418筛选阳性克隆,经RT-PCR与Western blot鉴定抑制效果,用MTT法观察抑制hif-1α基因表达后,HeLa细胞增殖的变化,RT-PCR检测在低氧培养条件下HeLa/shRNA-i细胞葡萄糖转运蛋白1mRNA的水平,利用MTT法观察抑制hif-1α表达后HeLa细胞对顺铂敏感性的改变。结果 HeLa/shRNA-i细胞与对照细胞组相比,hif-1α基因mRNA表达降低了62%,蛋白表达明显降低,MTT显示细胞吸光度值降低,低氧条件下葡萄糖转运蛋白1mRNA表达降低,对顺铂药物的敏感度增高,与对照组比较差异有统计学意义(P<0.05)。结论 hif-1α基因的沉默可以抑制HeLa细胞的增殖并且能够增强HeLa细胞对顺铂的敏感度。更多还原

【Abstract】 Objective To study the effects of silencing hif-1α gene expression on proliferation and influence on sensitivity of HeLa to cisplatin.Methods The expression vector of RNA interference(RNAi) system targeting hif-1α gene was constructed and transfected into HeLa cells with lipofectamineTM 2000.The positive clone was screened by G418,inhibition efficiency were measured with RT-PCR and Western blot.Glucose transport-1 mRNA was detected by RT-PCR,cell proliferation and sensitivity of HeLa to cisplatin were detected with MTT.Results Compared with control groups,HeLa/shRNA-i group showed decrease hif-1α expression at both at mRNA and protein levels.The mRNA level of glucose transport-1 was decreased.It was found that inhibition of hif-1α may increase drug sensitivity of HeLa cell to cisplatin.Conclusion Inhibition of the expression of hif-1α can reduce the proliferation and enhance the drug sensitivity of HeLa cells to cisplatin.更多还原