逆转录病毒载体介导的 HyTK 基因在人膀胱癌细胞株 EJ 中的表达

Retrovirus mediated HyTK gene transfer in human bladder cancinoma cell line EJ

为探索ＨＳＶ１ＴＫ基因对人膀胱癌的治疗作用，作者用一种与潮霉素磷酸转移酶（ｈｐｔ）基因相融和的Ⅰ型单纯疱疹病毒胸苷激酶（ＨＳＶ１ＴＫ）基因（简称ＨｙＴＫ基因）作目的基因，用逆转录病毒载体将ＨｙＴＫ基因导入人膀胱癌细胞株ＥＪ细胞中表达。含ＨｙＴＫ基因的逆转录病毒载体ＬＨｙＴＫＮ，经包装细胞ＰＡ３１７包装后，获得重组逆转录病毒；病毒感染人膀胱癌细胞株ＥＪ细胞，经潮霉素Ｂ筛选培养，获得潮霉素Ｂ的抗性克隆；ＰＣＲ扩增证实：病毒感染的ＥＪ细胞中导入了ＨＳＶ１ＴＫ基因，为开展ＨｙＴＫ基因治疗膀胱癌打下了基础。

HyTK gene,a fusion gene encoding both hygromycin phosphotransferase (hpt)and herpes simplex virus type 1 thymidine kinase (KSV 1 TK),was transduced into human bladder cancinoma cell line EJ by using recombinant retroviral vector.The recombinant retroviral vector LHyTKN containing HyTK gene was transduced into packaging cell line PA317.Recombinant retroviral supermant was obtained and employed to infect human bladder cancinoma cell line EJ cells.After Hygromycin B selection,hygromycion B resistant colonies were obtained and HSV 1 TK gene could then be detected in the retrovirus infected EJ cells by polymerase chain reaction.This laid the foundation to develop HyTK gene therapy for human bladder cancinoma.