顺铂联合胞嘧啶脱氨酶自杀基因对直肠癌细胞杀伤作用的研究

目的探讨顺铂联合胞嘧啶脱氨酶(CD)自杀基因对直肠腺癌细胞8348的杀伤作用。方法以绿色荧光蛋白(GFP)基因作为报告基因,观察在顺铂作用下GFP基因对8348细胞的转染效率;用克隆形成试验观察顺铂对CD基因转染效率的影响;同时用四甲基偶氮唑盐(MTT)试验检测顺铂联合CD/5-氟胞嘧啶(5-FC)对8348细胞的杀伤作用。结果顺铂提高质粒对8348细胞的转染效率26倍。对腺癌细胞的杀伤效率:单纯转染CD基因组为14.9%;IC50的氟尿嘧啶(5-FU)联合CD基因组为54.9%;IC50的顺铂联合CD基因组为88.5%,与其他两组相比,对癌细胞杀伤效率明显增强(P<0.01)。结论顺铂联合CD自杀基因能更有效地杀伤直肠腺癌细胞,可成为治疗直肠癌术后复发及转移的一种有效的方法。

Cisplatin combined with cytosine deaminase suicide gene therapy enhances the killing of rectal carcinoma cells in vitro

Objective To investigate the effect of cisplatin on gene transfer and the antitumoral effect of cisplatin combined with cytosine deaminase(CD) gene therapy on rectal carcinoma cells. Methods Green fluorescent protein(GFP) could be seen under fluorescent microscope.GFP gene were used for reporting gene to learn gene transfer efficiency under the influence of cisplatin.G418 were used to select CD positive neoplasm cells and CD gene transfer efficiency were tested by cloning efficiency. Antitumoral effect of cisplatin combined with CD/5 FC on rectal carcinoma cells were examined by MTT. Results Cisplatin could increase plasmid DNA transfer efficiency for 26 times. IC50 cisplatin combined with CD/5 FC could kill 88 percents of rectal carcinoma cells,while in the control group,5 FC only killed 15 percents of rectal carcinoma cells which were transduced with CD gene. Conclusion Cisplatin combined with CD suicide gene therapy may be used as a promising method for treating rectal neoplasm.