bcl-2基因转染小鼠造血干细胞重建造血的生物学特性  

Biological properties of reconstructed mouse hematopoiesis with hematopoietic stem cells transfected by bcl 2 gene.

作　　者：赵钧铭[1] 褚建新[1] 丁顺利[1] 姜露[1] 隋雪梅[1] 王淑萍[1] 

机构地区：[1]中国医学科学院中国协和医科大学血液学研究所

出　　处：《中华医学杂志》1999年第1期50-53,共4页National Medical Journal of China

基　　金：国家自然科学基金

摘　　要：目的观察ｂｃｌ２基因转染小鼠造血干细胞重建造血的小鼠对放化疗的抵抗作用。方法（１）含ｂｃｌ２ｃＤＮＡ的逆转录病毒载体质粒，经脂质体介导转染ＰＡ３１７包装细胞，通过Ｇ４１８筛选和克隆，获得高病毒滴度的细胞株（Ｄ３）；（２）用Ｄ３细胞株的病毒上清转染小鼠胚胎肝造血干细胞；（３）给致死剂量照射小鼠输注ｂｃｌ２转染的造血干细胞重建造血；（４）采用脾集落形成和再增殖能力观察ｂｃｌ２转染造血干细胞重建造血小鼠对化疗和放射的抵抗作用。结果ｂｃｌ２转染的造血干细胞的ＢＣＬ２蛋白表达率增高２５％，对照７％，其重建造血小鼠的存活率（７９％）高于对照组（５１％），白细胞恢复过程与对照组无明显差别；重建造血的动物可抵抗大剂量联合化疗和中等剂量放射造成的损伤，脾集落形成能力和脾集落再增殖能力均明显高于对照组；重建造血的动物活存一年以上，未见肿瘤发生。结论ｂｃｌ２基因转染造血干细胞重建造血有抗放化疗作用；即使只有部分造血干细胞转染ｂｃｌ２基因。Objective To observe resistance to radiochemical agents in the mice reconstructed hematopoiesis with hematopoietic stem cells transfected by bcl 2 gene.Methods The plasmid containing bcl 2 cDNA retrovirus vector was transfected to PA317 cells with Lipofecaine TM kite. A PA317 cell clone(D3) with high virus titer was obtained through G418 selection. The virus supernatant of D3 clone cells was used to transfect mouse embryo liver hematopoietic stem cells. Hematopoiesis in the mice treated with lethal dose irradiation was reconstructed by hematopoietic stem cells transfected with bcl 2 gene. The resistance to radiochemical agents in the mice with reconstructed hematopoiesis was evaluated according to the number of WBC of periphereal blood and mononuclear cells of bone marrow, CFU S and CFU S regeneration. Results The expression of bcl 2 protein was increased in the hematopoietic cells transfected by bcl 2 gene.The mice reconstructed hematopoiesis by stem cells transfected bcl 2 gene can tolerate high dose combined with chemotherapy and radiation of middle dose. The mice reconstructed hematopoiesis with stem cells transfected bcl 2 gene survived over one year, no tumors were found.Conclusion The mice with reconstructed hematopoiesis by stem cells carried bcl 2 gene possess the ability of resistance to radiochemical agents, even only part of hematopoietic stem cells were transfected with bcl 2 gene.

关 键 词：白血病 造血干细胞 抗药性 重建造血 转基因动物 

分 类 号：R733.705[医药卫生—肿瘤] R73－361[医药卫生—临床医学]