携带尤文肉瘤EWS-FLI-1结合序列的DTA表达载体构建及其对尤文肉瘤细胞杀伤作用的研究  

Construction of an Ewing's sarcoma specific expression vector of diphtheria toxin and observation of the killing effect on cultured Ewing's sarcoma cell line

作　　者：冯岗[1] 赵忠全[1] 王东林[1] 

机构地区：[1]第三军医大学新桥医院肿瘤科

出　　处：《第三军医大学学报》2004年第5期382-385,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 ( 30 10 0 189)~~

摘　　要：目的　研究尤文氏肉瘤EWS FLI 1融合蛋白的特异结合序列与DTA基因融合后的表达及对尤文肉瘤细胞的杀伤作用 ,为肿瘤基因治疗探索新的治疗方法。方法　构建含有EWS FLI 1特异结合序列和白喉毒素A链基因的表达载体pS2 DTA。转染尤文肉瘤细胞以及对照细胞后检测DTA表达及其对细胞的杀伤作用。结果　表达载体pS2 DTA在尤文肉瘤细胞系中高表达并产生强烈的杀伤作用。结论　pS2 DTA可以对尤文肉瘤细胞产生选择性杀伤作用 ,抑制其生长 。Objective To construct an expression vector containing Ewing's sarcoma EWS FLI 1 specific binding sequence and diphtheria toxin A chain (DTA) gene sequence to investigate the killing effect of DTA on cultured Ewing's sarcoma cells so as to provide proof for the exploration of new methods for gene therapy for cancer. Methods Expression vector (pS2 DTA) containing EWS FLI 1 specific binding sequence and DTA gene sequence was constructed by replacing the luciferase gene in pS2 with DTA gene at the points of Nco Ⅰ and Xba Ⅰ by molecular technique. After transfection of pS2 DTA into Ewing's sarcoma cells and the control cells, DTA expression and its killing effect on cells were detected. Results pS2 DTA was highly expressed in Ewing's sarcoma cell line, and the killing effect of DTA was much higher than that in the control cells. Conclusion pS2 DTA has selective killing effect on Ewing's sarcoma cells and can inhibit the growth of Ewing's sarcoma cells. So pS2-DTA has potential value in the treatment of Ewing's sarcoma.

关 键 词：尤文肉瘤 融合基因 白喉毒素A链(DTA) 基因治疗 

分 类 号：Q781[生物学—分子生物学] R394-33[医药卫生—医学遗传学]