Fas基因转染联合顺铂对直肠癌细胞杀伤作用的研究  

The effect of Fas gene transfection in combination with cisplatin on rectal carcinoma cells in vitro

作　　者：魏家臣[1] 李世拥[1] 安平[1] 于波[1] 蔡慧芸[1] 

机构地区：[1]北京军区总医院普通外科,100700

出　　处：《中华普通外科杂志》2003年第8期485-487,共3页Chinese Journal of General Surgery

基　　金：军队十五医学重点基金资助项目 ( 0 1Z0 0 6)

摘　　要：目的探讨Fas基因转染联合顺铂对直肠癌细胞的杀伤作用。方法采用RT PCR技术从健康人的外周血单核细胞中扩增包含全部阅读框架的Fas全长基因 ,与 pGEM TEasy质粒连接 ,经测序验证。构建 pcDNA3 1 Fas真核表达载体 ,将人Fas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基因mRNA表达。用MTT法分析顺铂对转染前后直肠癌细胞抑制增殖和诱导凋亡的作用。结果Fas基因转染可明显增强直肠癌 8348细胞的Fas表达 ,分别用 1、5、10、2 0、4 0mg/L浓度的顺铂作用 ,转染Fas基因的 8348细胞实验组细胞抑制率分别为 4 7 2 %、5 1 8%、5 7 2 %、6 5 4 %、71 0 % ;未转染Fas基因的 8348细胞对照组细胞抑制率分别为 2 9 6 %、33 0 %、37 8%、4 1 4 %、4 7 0 % ,2组相比 ,差异有显著性意义 (t =15 33,P <0 0 1)。结论转染的Fas基因可显著上调直肠癌 8348细胞的Fas表达 ,促进细胞凋亡 ,Fas基因转染联合顺铂对直肠癌细胞有更强的杀伤作用。Objective To study the effect of Fas gene transfection on rectal carcinoma cells in vitro. Methods By using RT-PCR technique, a full length of Fas gene 1007 bp was cloned from actived peripheral mononuclear cells of healthy donors. The fragment was ligated with the pGEM-T Easy and sequenced. The constructed vector was transfected into 8348 cells with lipofectin, the change in expression of Fas gene was determined by RT-PCR. The apoptosis and proliferation of rectal carcinoma cells pre- and posttransfection induced by cisplatin were analysed by ladder and MTT methods. Results Transfection of Fas gene significantly upregulates the expression of Fas in human rectal carcinoma 8348 cells. With the concentration of cisplatin at the level of 1, 5, 10, 20 and 40 mg/L , respectively, the suppression rates of Fas transfection 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(t=15.33, P<0.01).Conclusion Transfection of Fas gene by lipofectin significantly upregulates the expression of Fas in target cells. A strong suppression of rectal carcinoma cells was observed by combined Fas gene transfection and cisplatin.

关 键 词：FAS 基因转染 顺铂 直肠癌细胞 杀伤作用 研究 

分 类 号：R735.37[医药卫生—肿瘤]