基因枪在肿瘤基因治疗中的应用价值  

The use of gene gun in cancer gene therapy

作　　者：查园园[1] 林晨[1] 梁萧[1] 付明[1] 张雪艳[1] 吴旻[1] 

机构地区：[1]中国医学科学院中国协和医科大学肿瘤研究所分子肿瘤学国家重点实验室,北京100021

出　　处：《中华医学杂志》2000年第7期522-525,共4页National Medical Journal of China

基　　金：国家"八六三"高科技计划资助项目!(z200102)

摘　　要：目的　研究基因枪经小鼠皮肤转移基因的效率、被转基因的表达量、表达时间 ,以及对小鼠肿瘤的治疗效果 ,探讨作为一种新的基因转移工具 ,基因枪在基因治疗中应用的可能性。方法　(1)用荧光显微镜观察基因枪对体外培养细胞系的基因转导效率和被转基因表达时间 ;(2 )通过基因枪向小鼠皮肤转Lac Z基因后 ,取转基因部位的皮肤做冰冻切片进行 β gal染色 ,观察转基因蛋白在皮肤中表达的部位 ;(3)用ELISA法检测向小鼠皮肤转mGM CSF基因后 ,血清中的转基因蛋白表达量 ,并取转基因部位的皮肤做病理切片 ,通过观察局部病理改变 ,推测转基因蛋白在局部的表达情况 ;(4 )用基因枪向小鼠腹部皮肤转mGM CSF 10d后 ,用肿瘤细胞攻击 ,观察肿瘤生长情况 ;(5 )在小鼠右腋下接种肿瘤细胞 ,1周后进行转基因或注射瘤苗治疗 ,观察肿瘤生长情况。结果　 (1)基因枪能有效地对体外培养的各种肿瘤细胞系转基因 ,转导基因的效率是 16 .5 %± 9.0 % ,被转基因可获得较长时间的瞬时表达 ;(2 )小鼠皮肤冰冻切片染色结果显示 ,基因枪可有效地将外源基因导入到皮肤表皮 4～ 5层 ;(3)ELISA结果显示 ,经皮肤转基因后 ,血清中GM CSF蛋白水平显著升高 ,最高可达对照组的12 .5倍。小鼠皮肤病理切片可见到炎性浸润 。Objective With Helios gene gun ,the report genes EGFP and Lac Z were transfected to several cultured mammalian tumor cell lines in vitro and mice skin in vivo, respectively. A stable long time exogene's expression were got. Then , the pWRG3142, which carried mGM CSF gene was delivered to the abdominal skin of C57BL/6 mice using gene gun. Pathological sections showed the local transproteins' expression companying with a profound inflammation reaction characterized by neutrophilic infiltration . ELISA assay of transfected mouse's serum sample indicated a high improvement of transgenic proteins level , which demonstrated that transgenic GM CSF secreted from treated skin into the bloodstream effectively . In B16 melanoma tumor model , mice immunized with mGM CSF expression plasmids could be partially protected from 1×10 5 B16 cells challenge and exhibited a drastically reduced tumor growth rate . Finally, we conclucled that exogenes could be transfected into cultured cell lines and mice skin effectively by gene gun technology , and gene gun mediated in vivo delivery of GM CSF cDNA should be further developed for potential clinical testing as an approach for human cancer gene therapy.

关 键 词：生物弹道学 肿瘤 基因治疗 基因枪 

分 类 号：R730.5[医药卫生—肿瘤]