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转染金属硫蛋白基因1A促进大鼠肝星状细胞增殖
作者:王科  陈平圣 
单位:东南大学基础医学院 病理学与病理生理学系, 江苏 南京 210009
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; R-33; Q786
出版年·卷·期(页码):2009·28·第三期(189-193)
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.

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