GFP共表达的重构型Caspase-3基因对Hep-2细胞的作用

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.

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

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 recombinant Caspase-3 was observed by RT-PCR. The cell morphological changes were observed under fluorescent and (electronic) microscope and the cell survival rate after transfection 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.