Fas/FasL基因转染联合顺铂对直肠癌细胞杀伤作用的研究

Effect of Fas/FasL gene transfer combined with cisplatin on rectal carcinoma cells in vitro

目的探讨Fas/FasL基因转染联合顺铂对直肠癌细胞的杀伤作用。方法构建pcDNA3.1-Fas/FasL真核表达载体,将人Fas/FasL基因通过脂质体导入直肠癌8348细胞中,并利用RT-PCR方法检测直肠癌8348细胞的Fas/FasL基因mRNA表达。用MTT法分析顺铂对转染前后的8348细胞抑制增殖和诱导凋亡的能力。结果Fas/FasL基因转染可明显增强直肠癌8348细胞的Fas/FasL表达。分别加入不同浓度顺铂(1、5、10、20、40μg/ml),Fas转染组8348细胞抑制率分别为47.2%、51.8%、57.2%、65.4%、71.0%;后者细胞抑制率分别为29.6%、33.0%、37.8%、41.4%、47.0%,其差异有显著性意义(t=15.33,P<0.01);FasL转染组8348细胞抑制率分别为11.0%、25.4%、31.2%、37.8%、42.4%;对照组8348细胞抑制率分别为26.1%、34.4%、37.6%、42.9%、53.2%,其差异有显著性意义(t=4.43,P<0.05)。结论转染的Fas/FasL基因可显著上调直肠癌8348细胞的Fas/FasL表达;Fas基因转染联合顺铂对直肠癌细胞有更强的杀伤作用;FasL基因转染可减弱顺铂对8348细胞的细胞毒作用,因此为直肠癌的基因治疗和化疗提供了理论依据。

Objective To study the effect of Fas/FasL gene transfer combined with cisplatin on rectal carcinoma cells in vitro. Methods The pcDNA3.1 Fas/FasL vector was constructed and was transfected into 8348 cells with lipofectin. Fas/FasL gene expression was determined by RT PCR. Proliferation suppression and apoptosis of 8348 cells pre and post transfection induced by cisplatin were analysed by MTT methods.Results Transfer of Fas/FasL gene can significantly upregulated the expression of Fas/FasL in human rectal carcinoma 8348 cells. At different concentrations of cisplatin, the suppression rates of Fas transfer group and control group were 47.2%,51.8%,57.2%,65.4%,71.0% and 29.6%,33.0%,37.8%,41.4%,47.0% respectively,whose difference were all significant(t=15.33,P< 0.01). The suppression rates of FasL transfer group and control group were 11.0%,25.4%,31.2%,37.8%,42.4% and 26.1%,34.4%,37.6%,42.9%,53.2% respectively,whose differences were all significant(t=4.43,P< 0.05). Conclusions Fas gene transfer can enforce the killing effect of cisplatin on tumor cells. Combined use of FasL gene transfer and cisplatin may help in the treatment of cisplatin resistant rectal carcinoma, providing a basis for gene therapy and chemotherapy of rectal carcinoma.