转染CD258-Fc基因抑制人膀胱移行细胞癌T24细胞增殖的体内外实验研究  

Studies of CD258-Fc gene transfection into human bladder transitional cell carcinoma T24 cell lines inhibition proliferation in vivo and in vitro

作　　者：黄东龙[1] 张轶庠[1] 张辉 杨江根[1] 罗思颖 刘红艳[1] 

机构地区：[1]暨南大学医学院第二附属医院(广东省深圳市人民医院)泌尿外科,广东深圳518020

出　　处：《中国现代医学杂志》2007年第7期777-781,共5页China Journal of Modern Medicine

基　　金：深圳市科技计划项目(医药卫生类No.200603080)

摘　　要：目的探讨转染CD258-Fc基因抑制人膀胱移行细胞癌T24细胞增殖的作用及对人膀胱移行细胞癌动物模型抑瘤率的影响,以探讨CD258-Fc基因转染实现抗肿瘤的作用,为人膀胱移行细胞癌的基因治疗提供新思路。方法以LipofectaminTM脂质体介导CD258-Fc基因转染人膀胱移行细胞癌T24细胞株,通过绘制细胞生长曲线、MTT比色法以及流式细胞仪检测观察CD258-Fc转染对T24细胞增殖的影响,PCR、Northern分析检测转染CD258-Fc基因在T24细胞的表达,WesternBlot检测蛋白表达,T24细胞接种于裸鼠,分组给予CD258-Fc基因转染进行治疗,评价其抑瘤率。结果CD258-Fc基因的表达可以抑制T24细胞的体外生长,T24/CD258-Fc细胞的生长曲线较对照组明显降低;MTT比色显示T24/CD258-Fc的细胞活力与对照组相比有显著性差异(P<0.05);PCR及Northern分析提示CD258-Fc基因转染后在T24细胞上调表达,WesternBlot检测到蛋白高表达;流式细胞仪检测显示基因转染使瘤细胞的生长滞留在G0+G1期,S期细胞明显减少,并观察到细胞凋亡。动物实验T24/CD258-Fc、T24/CD258-Fc+IFN-γ肿瘤抑制率可达48.66%和60.98%,显著高于对照组(P<0.05),且两组间差异具有统计学意义(P=0.032)。结论CD258-Fc基因转染对T24细胞具有体内外抗肿瘤的作用,提示CD258-Fc基因转染T24细胞技术有可能为人BTCC的基因治疗提供一种新的治疗策略与手段。[Objective] To investigate the inhibition proliferation effect of CD258-Fc on human bladder transitional cell carcinoma T24 cell lines in vitro and in vitro and the inhibitory rate in nude mice model so as to discuss CD258-Fc gene transfection into T24 cell lines seeking a new way for gene therapy of carcinoma of human Bladder Transitional Cell Carcinoma （BTCC）. [Methods] CD258-Fc expression vector or pc DNA 3.1 （＋） blank vector was transfected into human BTCC T24 cell lines and normal T24 ceils were used as normal control. The inducing apoptosis effects of CD258-Fc gene on cell growth was described by cell growth curve,MTT assay and flow cytometry （FCM） analysis. The expression of CD258 gene after transfection was detected by RT-PCR, Western blot assay for the protein. The nude .mice model carried BTCC T24 cell lines were constructed, the nude mice were consecutively treated by CD258-Fc gene transfection, the inhibitory tumor rate was estimated. [Results] Expression of CD258-Fc gene strongly inhibited T24 cell proliferation in vitro. The growth of T24,CD258 was significantly lower than that of control. MTT test showed that the difference of cell viability between T24,CD258 and control cells was also significant （P 〈0.05）. The expression of CD258 gene after transfection was highly detected by RT-PCR, the high expression of protein was detected by Western blot assay. FCM showed T24/CD258 cells on S phase was reduced greatly and apoptosis could be observed in some cells. In nude mice of BTCC model, the inhibitory rate in CD258-Fc gene transfection group without and with different IFN-γ concentrations got up to 48.66% and 60.98% （P 〈0.05）, which differed remarkably （P =0.032）. [ Conclusions] The findings showed that the transfeetion of CD258-Fc gene induced cellular apoptosis in T24 cells and that the BTCC of the nude mice model were inhibited by CD258-Fc gene transfeetion which suggested that CD258-Fc gene transfeetion into T24 cells technique can be an effective means in the gene the

关 键 词：CD258-Fc 转染 T24细胞 基因治疗 

分 类 号：R730.5[医药卫生—肿瘤]