关键词: 天冬氨酸蛋白酶类;脱噬作用;肿瘤;基因疗法

    摘 要:目的 观察重构型caspase-3及其N-端融合蛋白的表达对HeLa细胞的凋亡诱导作用. 方法 用反转录PCR获取人caspase-3基因，用重组PCR方法构建人重构型caspase-3及其融合蛋白基因，将基因克隆入表达载体pcDNA3，脂质体法转染HeLa细胞，通过细胞计数、电镜观察、MTT法等检测被转染细胞的生长及其凋亡状况. 结果 成功获得了cas-pase-3基因，构建了重构型caspase-3基因和重构型caspase-3与绿脓杆菌外毒素(PE)转膜结构域部分肽段的融合蛋白基因及其表达载体，与对照组相比，在转染后1～4d内，两种基因的表达均导致HeLa细胞存活数减少，大量细胞发生凋亡，且二者活性相当. 结论 重构型caspase-3及其融合蛋白可以诱导转染的HeLa细胞发生凋亡.

    Construction and effect of caspase┐3fusion proteins and their apoptosis induction in HeLa cells

    JIA Lin-Tao，YU Cui-Juan，XU Yan-Ming，ZHU Feng，PENG Wei-Dan，SU Cheng-Zhi，WANG Cheng-Ji，YANG An-Gang

    Department of Biochemistry and Molecular Biology，Faculty of Preclinical Medicine，Fourth Military Medical University，Xi'an710033，China

    Keywords:aspartic proteinases;apoptosis;neoplasms;gene therapy

    Abstract:AIM To investigate the induction of HeLa cells to apoptosis by the expression of reversed caspase-3and its N-terminal fusion protein.METHODS RT-PCR and recombi-nant PCR were used to amplify human caspase-3gene and construct reversed caspase-3and its fusion protein genes.Following cloning into expression vector pcDNA3and trans-fecting HeLa cells ......