转染金属硫蛋白基因1A促进大鼠肝星状细胞增殖  

Metallothionein gene transfected enhancing the proliferation of hepatic stellate cells

作　　者：王科[1] 陈平圣[1] 

机构地区：[1]东南大学基础医学院病理学与病理生理学系,江苏南京210009

出　　处：《东南大学学报（医学版）》2009年第3期189-193,共5页Journal of Southeast University(Medical Science Edition)

基　　金：国家自然科学基金资助项目(30470780)

摘　　要：目的:观察金属硫蛋白基因1A(MT1A)对肝星状细胞(HSCs)增殖的影响。方法:构建MT1A基因真核表达质粒,转染HSCs。通过RT-PCR、Western blotting法检测导入的质粒在HSCs中的表达,采用AgNOR染色法及Ki-67抗原免疫细胞化学染色检测HSCs增殖指数。结果:MT1A基因在HSCs中过表达;转染MT1A基因组每个细胞AgNOR颗粒数(5.43±0.45)较pcDNA3.1组(3.94±0.39)及非转染组(4.14±0.34)均高(P<0.05);免疫细胞化学染色结果显示,转染MT1A基因组细胞Ki-67染色呈阳性,而对照组为阴性。结论:外源MT1A基因在HSCs中过表达可促进其增殖。Objective To investigate the effect of metallothionein 1A gene（MT1A） on the proliferation of rat hepatic stellate cells（HSCs） in vitro.Methods MT1A expressing plasmid（MT1A-pcDNA3.1） and pcDNA3.1 empty plasmid were transfected into HSCs by fuGENE-6 transfection reagent respectively.The expression of MT1A in HSCs was detected by RT-PCR and Western blotting,and the HSCs proliferation changes were assessed by AgNOR staining and immunocytochemistry staining of the Ki-67 antigen.Results It was found that MT1A mRNA and protein showed overexpression in the cells transfected MT1A-pcDNA3.1.AgNOR staining showed that the number of average particles in each nucleus was 5.43±0.45 in the cells overexpressing MT1A,whereas only 3.94±0.39,4.14±0.34 in the cells expressing the empty vector and untransfected cells respectively.Proliferation indexes measured with AgNOR staining in MT1A overexpression cells were significantly higher than that in the control cells（P〈0.05）.Ki 67 antigen presented mainly in the nucleus in MTs overexpression cells,and no positive staining was found in the control cells.Conclusion The results suggest that MT1A may enhance HSCs proliferation in vitro.

关 键 词：肝星状细胞 金属硫蛋白 增殖 肝纤维化 大鼠 

分 类 号：Q253[生物学—细胞生物学] R-33[医药卫生]