垂体腺瘤双重靶向性基因治疗的实验研究  

Experimental study of dually targeting gene therapy system for pituitary adenomas

作　　者：马驰原[1] 卢亦成[2] 史继新[1] 任常春[3] 朱景德[3] 顾健人[3] 

机构地区：[1]南京军区南京总医院神经外科,210002 [2]第二军医大学长征医院神经外科 [3]上海市肿瘤研究所癌基因及相关基因国家重点实验室

出　　处：《中华医学杂志》2005年第4期262-266,共5页National Medical Journal of China

摘　　要：目的构建并实验评价转染与转录双重靶向性基因治疗系统对垂体腺瘤的治疗作用。方法构建GE7基因导入系统介导的生长激素启动子调控的基因治疗系统,通过对垂体生长激素腺瘤GH3细胞的体外实验,以及垂体腺瘤裸鼠模型的体内实验,观察此系统用于垂体腺瘤基因治疗的可能性和靶向性。结果成功构建双重靶向性基因治疗系统;基因转染后,GH3细胞可检测到HSVTK蛋白,对照细胞为阴性;在此基础上予以GCV(4mg/L),GH3细胞存活率平均为106%,相同条件下,U2OS、HO8910PM的存活率分别为829%和935%(P<001);裸鼠皮下种植肿瘤经基因治疗后,治疗组肿瘤体积明显缩小(168mm3±17mm3),各对照组肿瘤体积均有不同程度的增大(P<001);治疗组裸鼠的生存期也较对照组明显延长(平均123d和40d,P<001)。结论GE7转染的生长激素启动子调控的基因治疗有望成为垂体腺瘤的靶向性治疗策略。Objective To construct a dually targeting gene therapy system for pituitary adenomas and investigate its effect. Methods Promoter hGHp containing human growth hormone gene was obtained from human genome and cloned into the plasmid pcDNA3.1/His A with the promoter cut to construct the recombinant plasmid pcDNA3.1/His A-hGHp. HSV-TK gene was obtained from the plasmid pcDNA3.1/His A-TK and integrated into the plasmid pcDNA3.1/His A-hGHp to construct the recombinant plasmid pcDNA3.1/His A-hGHp-TK. A GE7 gene delivery system-mediated human growth hormone promoter controlled gene therapy system was constructed by adding the mixture of GE7-polylysine and HA20-polylysine into the DNA solution. Human growth hormone-secreting pituitary adenoma cells of the GH_3 line, human myeloma cells of the U-2OS line, and human oophoroma cells of the HO8910PM line were cultured and transfected with PBS or GE7 packaged pcDNA3.1/HisA-TK or pcDNA3.1/HisA-hGHp-TK. Western blotting was used to examine the expression of PBS or GE7 packaged pcDNA3.1/HisA-TK or pcDNA3.1/HisA-hGHp-TK protein , MTT method was used to detect the cell survival rate. Another GH3, U-2OS, and HO8910PM cells were cultured and transfected with PBS or GE7 packaged pcDNA3.1/HisA-TK or pcDNA3.1/HisA-hGHp-TK and then ganciclovir (GCV) was added. MTT method was used to examine the cell survival rates. GH3 cells were injected subcutaneously into the right axilla of 200 SD nude rats loaded with human pituitary adenoma. Three weeks after the rats were randomly divided into 5 equal groups: PBS group in which PBS was injected into the tumor and GCV was injected peritoneally; GE7 group in which GE7-polylysine and HA20-polylysine were injected into the tumor and GCV was injected peritoneally; without TK group in which GE7-packaged pcDNA3.1/His A-hGHp was injected into the tumor and GCV was injected peritoneally; without GCV group in which GE7-packaged pcDNA3.1/His A-hGHp-TK was injected into the tumor and PBS was injected peritoneally; and treatment group in which GE7-packaged pcD

关 键 词：垂体腺瘤 基因治疗 靶向性 GH 治疗系统 对照组 性基因 基因导入 下种 种植 

分 类 号：R736.4[医药卫生—肿瘤] R739[医药卫生—临床医学]