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目的探讨上下调CD59锚固蛋白对Hela细胞增殖活性的影响。方法将前期构建的pSUPER-siCD59质粒、pIRES-WT CD59质粒以及增加了CD59W40活性位点(C39W40K41→W39W40W41)的pIRES-MCD59质粒用脂质体转染的方法转染Hela细胞,免疫荧光法倒置荧光显微镜下观察Hela细胞膜上CD59基因表达,并用RT-PCR及Western blot的方法检测各组转染细胞中CD59的表达。采用噻唑蓝(MTT)比色法检测各转染组CD59抗体交联前后Hela细胞增殖活性的变化,比较各组差异。结果转染pSUPER-siCD59质粒组CD59基因与蛋白表达量明显低于正常组,细胞增殖活性降低,而转染pIRES-WT CD59与pIRES-MCD59质粒组则反之。结论下调CD59能抑制Hela细胞增殖活性,上调CD59则能增强其增殖,为宫颈癌的靶向治疗奠定了基础。
Objective To investigate the effect of down-regulation of CD59 anchoring protein on the proliferation of Hela cells. Methods Plasmid pSUPER-siCD59, pIRES-WT CD59 plasmid and pIRES-MCD59 plasmid with increased CD59W40 active site (C39W40K41 → W39W40W41) were transfected into Hela cells by lipofection. The immunofluorescence inverted fluorescence The expression of CD59 on Hela cell membrane was observed under a microscope. The expression of CD59 in transfected cells was detected by RT-PCR and Western blot. The proliferation of Hela cells was detected by MTT assay before and after cross-linking of CD59 antibody. The differences among groups were compared. Results The expression of CD59 gene and protein in pSUPER-siCD59 plasmid group was significantly lower than that in normal group, while the cell proliferation activity was decreased. However, the transfection of pIRES-WT CD59 and pIRES-MCD59 plasmid group was the opposite. Conclusion The down-regulation of CD59 can inhibit the proliferation of Hela cells. Upregulation of CD59 can enhance the proliferation of Hela cells and lay a foundation for the targeted therapy of cervical cancer.