Fas基因转导大肠癌细胞对化疗药物作用敏感性的实验研究

Sensitivity of the Fas-expressed Colorectal Cancer Cells to Chemotherapy

目的:观察表达Fas基因的大肠癌细胞株对化疗药物的敏感性,为今后大肠癌的基因治疗、临床化疗打下实验基础。方法:应用四甲基偶氮唑盐(MTT)微量酶反应观察Fas转导细胞株(Lovo Fas cells)对化疗药物作用的敏感性,实验分组包括:非转导细胞组;②Fas转导细胞组;③Fas转导细胞+Fas抗体组(抗体结合组),Fas抗体浓度1∶100;各组分别培养3个复孔相应细胞株为对照组,未加化疗药物。将转导细胞和非转导细胞接种于96孔细胞培养板,各组分别加入顺铂(CDDP)、氟尿嘧啶(5-FU)、长春新碱(VCR)已知临床血浆高峰浓度的0.1倍、1.0倍及10倍的剂量,各设3个复孔,培养3天,加入MTT溶液,继续孵育4h,弃上清,加入二甲基亚砜(DMSO),酶联免疫检测仪上选择590nm波长测定各孔OD值。结果:各组细胞存活率比较,Fas转导细胞经过CD-DP、5-FU、VCR作用细胞存活率在0.1倍、1.0倍、10倍剂量均低于非转导细胞,但无统计学意义(P>0.05);抗体结合组经上述化疗作用,细胞存活率明显降低,与非转导细胞组和Fas转导细胞组比较,差异有非常显著性意义(P<0.01)。各组与未加化疗药物组比较,细胞存活率随药物浓度增加而明显下降(P<0.01),以抗体结合组最为明显。结论:Fas转导细胞株与Fas抗体联合作用较非转导细胞株对化疗药物的敏感性明显增强。

Objective：To observe the sentivity of Fas-expressed colorectal cancer cells to chemotherapy and to provide an experimental data for gene therapy and clinical chemotherapy. Methods： The sensitivity of Fas-expressed colorectal cancer cells to chemotherapy was observed by MTT. The groups were divided as： a. non-transfecting cells, b. Fas-transfecting cells and c. Fas-transfecting cells ＋ Fas antibody. The concentration of the Fas antibody was 1 ： 100. Fas-transfecting cells and non-Fas-transfecting cells were inoculated in the 96 hole cell culture band. Each group was added with Cisplatin （CDDP）, Fluorouracil （5-FU）and Vincristine （VCR）at the dose of 0.1,1.0 and 10 times of their plasma peak concentrations,respectively. Triplication was performed and cultured for 3 days and then MTT was added ,incubated for another 4 hours, DMSO was added before ELISA detection and,finally, OD index was obtained. Results ：The survival rate of Fas-transfected cells treated with CDDP,5-FU,VCR was lower than that of non-transfected cells at all concentrations,but the discrepancy was not significant. There was significant difference between the Fas-transfected cells ＋ Fas antibody group of non-transfected cells and Fas-transfected cells（P 〈 0. 01 ）. The survival rate of the former was much lower. It was suggested that the sensitivity of Fas-transfected cells to chemotherapy was enhanced when combined with Fas antibody. Conclusion：Our in vitro experiment shows that Fas-transfected cells combined with Fas-antibody are more sensitive to chemotherapeutic agents than those of non-transfected cells.