摘要
目的研究外源性线粒体融合素基因-2(mitofusin-2 gene,mfn2)对肝癌细胞Caspase蛋白表达的影响及意义。方法用脂质体2000将质粒pEGFPmfn2、空质粒pEGFP-N2分别转染肝癌细胞株HepG2,分别检测转染后48h HepG2细胞mfn2 mRNA的表达、蛋白表达及凋亡特异性蛋白酶Caspase3、Caspase 9及proCaspase 9蛋白表达的变化。结果经脂质体2000转染后,HepG2细胞基因组中存在有目的基因特异性片段,转染后细胞中有Mfn2蛋白表达;转染pEGFPmfn2组细胞中Caspase 3、Caspase 9表达明显增加(P<0.05),而proCaspase 9的表达明显减少(P<0.05)。结论外源性mfn2基因转染可激活Caspase 3、Caspase 9,进而激活细胞凋亡信号通路,这可能是mfn2基因诱导肝癌细胞凋亡的可能机制。
Objective To investigate the effect and significance of exogenous mitofusin-2 gene （mfn2） on expression of Caspase of hepatocellular carcinoma cells. Methods Plasmid pEGFPmfn2 and pEGFP-N2 were transfected respectively into hepatocellular carcinoma cell strain HepG2 by Lipofectamine^TM 2000 in vitro. The mRNA expression, protein expression and the expression of apoptosis related protein expression of Caspase 3, Caspase 9 and proCaspase 9 were detected. Results After transfected by Lipofectamin^TM 2000, the genome of mfn2-transfected cells contained a 392bp specific fragment of mfn2 gene which indicated the succeessful transfection, and successful protein expression was confirmed by Western Blot assay. The protein expression of Caspase 3, Caspase 9 were higher and procaspase 9 were lower in mfn2 transfected cells than in control group cells （P 〈 0. 05 ） . Conclusion Mfn2 could activate Caspase 3 and Caspase 9 which might contribute to explore the mechanism of its inducing apoptosis of hepatocellular carcinoma ceils.
出处
《广州医药》
2009年第4期3-5,共3页
Guangzhou Medical Journal
