摘要
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。
The purpose of this study is to observe the changes of Hela cells proliferation induced by up or down regulating of the anchoring protein CD59. By using Lipofectamin 2000, Hela cells were transfected with recombinant pSUPER-siCD59 plasmid, plRES-WTCD59 plasmid, and pIRES-MCD59 plasmid containing CD59＇s active site W40. Then immunofluorescence, RT-PCR and Western blot were carried out for the detection of CD59 expression both on RNA and protein levels of Hela cells. Subsequently, MTT colorimetry was employed to evaluate the proliferation activity of every Hela cell group. We found that Hela cells transfected with pSUPER-siCD59 plasmid demonstrated lower CD59 expression and lower cell proliferation compared with the normal groups. On the contrary, the groups transfected with pIRES-WTCD59 and plRES-MCD59 plasmids had the opposite effects. The results indicated that down-regulation of CD59 could restrain the proliferation activity of Hela cells, while up- regulation could strengthen it. The study should lay a foundation for the targeted therapy of cervical cancer.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2013年第11期943-947,共5页
Immunological Journal
基金
青岛市科技局项目(12-1-3-5-(2)-hsh)
关键词
CD59
上调
下调
HELA细胞
增殖
CD59
Up-regulation
Down-regulation
Hela cell
Proliferation
