摘要
目的构建恶性人脑胶质瘤细胞株TJ899的单纯疱疹病毒的胸腺激酶/丙氧鸟苷（HSV-Tk/GCV）自杀基因治疗系统，探讨该体系在脑胶质瘤治疗中的作用。方法采用PCR、RT-PCR、融合PCR、酶切、连接等技术构建由自杀基因pcDNA3.1（-）CMVTK表达载体；用脂质体介导法将pCMVTK质粒转染进入TJ899恶性人脑胶质瘤细胞，并用M IT法检测感染后细胞对（GCV）的敏感性。结果pCMVTK质粒经测序证实含有TK目的基因；脂质体介导Pcmvtk质粒成功转染了TJ899细胞，并在体外实验研究中显示很好的治疗效果。结论建立了作用于恶性人脑胶质瘤细胞株TJ899的HSV-Tk/GCV自杀基因治疗系统，为脑胶质瘤的临床研究打下了基础。
Objective To construct plasmid vector pcDNA3, 1 （ - ）CMVTK and evaluate the effects of Lipo-mediated in vitro gene transfer of pcDNA3. 1 （ - ） CMVTK in cell proliferation and anglo- genesis of malignant glioma cells TJ899, Methods We constructed the expression plasmid vector of pcDNA3, 1（ -）CMVTK with PCR, RT- PCR, confusion PCR, enzyme restriction, ligation and so on. Combined the Lipofection with suicide gene pcDNA3, 1 （ - ） CMVTK for transfection the TJ899 cells, MTF analysis was used to detect the cytotoxic effects of the pcDNA3, 1 （ - ） CMVTK/5-FC system, Result The plasmid vector contained the target TK gene which was confirmed by the sequencing, The Lipo-mediated pcDNA3, 1 （ - ） CMVTK were effectively transfered into TJ899 cells and showed high effective gene therapy in vitro, Conclusion We constructed plasmid vector pcDNA3, 1 （ - ） CMVTK gene therapy systems, which set up foundation of clinic therapy for malignant glioma.
出处
《中国医师杂志》
CAS
2007年第8期1037-1039,共3页
Journal of Chinese Physician
