FL转基因细胞的构建及膜型FL对U937细胞的体外促增殖效应  

Construction of the transfect cell line expressing the human FL gene and its biological effects to U937 cells in vitro

作　　者：居颂光[1] 居颂文[1] 傅晋翔[1] 仇红霞[1] 王明元[1] 周斌[1] 王凤鸣[1] 刘彤[1] 刘高勤[1] 邱玉华[1] 张学光[1] 

机构地区：[1]苏州大学医学院生物技术研究所,215000

出　　处：《中国现代医药杂志》2005年第5期1-4,共4页Modern Medicine Journal of China

基　　金：江苏省放射医学与防护重点实验室开放经费资助项目(KJS05031);国家自然基金青年科学基金项目(30400394)

摘　　要：目的构建稳定表达人膜型FL分子的转基因细胞株,观察膜型FL和可溶性FL对单核细胞性(M5型)白血病细胞株的体外促增殖效应。方法利用PCR方法克隆人FL全长基因,继而重组入逆转录病毒表达载体pEGZ-Term,转染包装细胞293T,用含有完整重组逆转录病毒颗粒的293T细胞培养上清感染L929细胞,筛选获得Zeocin抗性的转基因细胞。采用RT-PCR和流式细胞仪表型分析等方法进行鉴定。MTT法分析了L929/FL细胞和可溶性FL体外刺激U937细胞的增殖效应。结果成功构建了稳定表达人膜型FL的转基因细胞L929/FL,膜型FL在体外能够有效促进U937细胞增殖,而可溶性FL并不能有效促进U937细胞的体外增殖。结论成功构建了稳定表达人膜型FL的转基因细胞,并且提示膜型FL在白血病发病机理中可能具有重要作用。Objective To construct transgenie cell line that stably expressing human membrane FL and observed the proliferation effects of membrane and soluble FL to monoeytie leukemia cell line in vitro. Methods Full length FL gene was cloned by PCR and subcloned into retrovirus expression vector pEGZ-Term, the recombinant vector pEGZ/FL was transfected into the package cell 293T. The supernatant of 293T cells was used to infect L929 cells. Target protein stably expressed by L929 cells was analyzed by FCM. Then L929/FL transgenie cells and soluble FL were used to stimulate the U937 cells to observe their biological effects by MTT method. Results Human FL transgenic cell line L929/FL that stably expressing membrane was established and the membrane FL could promote the proliferation of U937. Conclusion Transgenic cell line stably expressing membrane FL was established. It suggested that membrane FL may play important roles in leukemia pathogenesis.

关 键 词：FL 转基因细胞 白血病 细胞株 U937细胞 转基因细胞 促增殖效应 FL细胞 体外刺激 膜型 逆转录病毒表达载体 白血病细胞株 L929细胞 

分 类 号：R733.7[医药卫生—肿瘤] R394[医药卫生—临床医学]