GFP共表达的重构型Caspase-3基因对Hep-2细胞的作用  

Effects of recombinant Caspase-3 on human laryngocarcinoma cell line Hep-2 through coexpression with GFP

作　　者：余晓林[1] 刘红[1] 陈万平[1] 程鹏远[1] 

机构地区：[1]第四军医大学门诊部,陕西西安710033

出　　处：《第四军医大学学报》2005年第21期1945-1947,共3页Journal of the Fourth Military Medical University

摘　　要：目的:探讨GFP共表达的Caspase-3基因的表达对人喉癌细胞Hep-2的凋亡诱导作用.方法:应用DNA重组技术将重构型Caspase-3基因亚克隆至带有GFP报告基因的真核表达载体pEGFP-C1中,脂质体介导转染人喉癌细胞Hep-2,通过RT-PCR方法检测转染后Caspase-3基因的表达;荧光显微镜、电镜观察细胞形态学变化;MTT法检测转染Caspase-3基因对Hep-2细胞生长的影响.结果:RT-PCR方法证实重构型Caspase-3基因可在Hep-2细胞内表达,形态学观察显示转染组细胞较对照组出现明显细胞凋亡特征,MTT法结果表明Caspase-3对Hep-2的细胞生长有抑制作用.结论:重构型caspase-3对喉癌细胞Hep-2的生长有抑制作用,并可诱导成骨肉瘤细胞凋亡.AIM： To investigate the apoptosis induction effect of the expression of recombinant Caspase-3 on laryngocarcinoma cell line Hep-2. METHODS： Recombinant Caspase-3 gene was subcloned into the GFP reporting vector pEGFP-C1 to generate the expressing vector pEGFP- Caspase-3 by DNA recombinant technique. Human laryngocarcinoma cell line Hep-2 was transfected with Caspase-3 gene by Lipofectamine 2000 and the expression of - nant Caspase-3 was observed by RT-PCR. The cell morphological changes were observed under fluorescent and electronic microscope and the cell survival rate after trans- fection was assayed by MTT method. RESULTS： Recombinant Caspase-3 gene was stably expressed in Hep-2. Recombinant Caspase-3 induced obvious apoptosis in vitro. The MTT results suggested that the cell curve declined after transfection with Caspase-3. CONCLUSION： Recombinant Caspase-3 can inhibit the growth of the laryngocarcinoma cell line Hep-2 and effectively induce it to apoptosis.

关 键 词：CASPASE-3基因 凋亡 基因转染 

分 类 号：Q255[生物学—细胞生物学]