pemt2-cDNA的转染抑制大鼠CBRH-7919肝癌细胞c-Met及Bcl-2的表达并诱发凋亡

Transfection of pemt2-cDNA Inhibits c-Met and Bcl-2 Expression and Induces Apoptosis in Rat CBRH-7919 Hepatoma Cells

为探讨磷脂酰乙醇胺 N 甲基转移酶 2 (phosphatidylethanolamine N methyltransferase 2 ,PEMT2 )的转染抑制大鼠肝癌CBRH 7919细胞增殖的分子机理 ,构建了带有完整的pemt2 cDNA基因质粒载体 ,经转染和鉴定 ,确知其在大鼠肝癌细胞系CBRH 7919细胞中稳定高表达 .用细胞培养、免疫细胞化学、Western印迹、流式细胞仪和琼脂糖凝胶电泳等技术研究c Met(HGF的受体 )及抗凋亡蛋白Bcl 2在此过程中的表达和是否诱发凋亡 .免疫组织化学及Western印迹分析显示在转染高表达细胞中 ,c Met及Bcl 2的表达下调 .流式细胞仪分析表明 ,在转染pemt2 cDNA的高表达细胞中 ,G1期细胞增加 ,S期细胞减少 ,并在G0 期出现一个凋亡峰 .琼脂糖凝胶电泳谱显示梯状条带 .结果表明 :pemt2的转染可使大鼠肝癌细胞的c Met及Bcl 2的表达下调 ,并发生凋亡 .

To unravel the molecular mechanism of inhibition of proliferation induced by transfection of pemt 2 cDNA into CBRH 7919 hepatoma cells,cell culture, pemt 2 cDNA transfection,flow cytometry,immunocytochemical and Western blotting,and agarose gel electrophoretic techniques were used.The expression of the HGF receptor c Met and the antiapoptotic protein Bcl 2 were down regulated in the pemt 2 transfected cells.The high expression clone of pemt 2 transfected cells showed an increase in cell number of G 1 phase and a decrease in that of S phase.An apoptotic peak was seen in the G 0 phase.DNA ladder was seen in agarose gel electrophoresis.The results show that transfection of pemt 2 into rat hepatoma cells could inhibit the expression of c Met and Bcl 2 and induce apoptosis.This could,at least partly,account for the reduced proliferation of the pemt 2 transfected CBRH 7919 cells.