基因枪介导的neu基因DNA疫苗抗肿瘤作用的实验研究  

Antitumor effect of gene gun-mediated DNA vaccine pWRG-neu immunization in C57BL/6 mice

作　　者：查园园[1] 林晨[1] 张叔人[1] 梁肖[1] 张雪艳[1] 付明[1] 张自强[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医院肿瘤研究所分子肿瘤学国家重点实验室,北京100021

出　　处：《中华肿瘤杂志》2003年第4期320-324,共5页Chinese Journal of Oncology

基　　金：国家高技术研究发展计划 ( 863计划 )课题( 2 0 0 1AA2 1710 1)

摘　　要：目的　探讨DNA疫苗pWRG neu的皮内免疫 ,对高表达neu基因的小鼠移植瘤生长和转移的抑制作用。方法　向小鼠黑色素瘤B16F10细胞系转染pcDNA neu ,用有限稀释法筛选一株高表达neu基因的细胞株B16F10 neu。在基因枪介导下 ,向C5 7BL/ 6小鼠导入DNA疫苗pWRG neu ,通过观察免疫动物的生存期 ,评价DNA疫苗的抗肿瘤作用。分离免疫动物脾细胞 ,经自体淋巴细胞混合培养实验 ,分析DNA疫苗体内免疫后机体的CTL应答。结果　筛选到一株高表达neu基因的B16F10 neu细胞株 ,转基因过程和外源基因的表达没有改变细胞系的增殖特性。用基因枪轰击 ,进行DNA疫苗pWRG neu皮内免疫 ,对小鼠黑色素瘤B16F10 neu进行预防、治疗和抗转移的实验研究 ,结果表明 ,DNA疫苗的免疫能够明显推迟移植瘤的生长 ,延长小鼠生存期 ,获得明显的抗肿瘤效果。DNA疫苗免疫后可诱导小鼠脾淋巴细胞CTL活性。结论　基因枪介导的DNA疫苗pWRG neu经皮内免疫 ,能够有效的诱导机体的细胞免疫应答 ,预防和治疗小鼠移植瘤的发生 ,并有一定的预防肿瘤肺转移的作用。Objective To investigate the inhibitory effect of DNA vaccine immunization on neu overexpressed melanoma growth in prophylactic treatment and anti lung metastasis experiments in C57BL/6 mice. Methods pcDNA neu transfected into B16F10 with transfection reagent Fugene TM 6, neu overexpressed cell clone B16F10 neu was selected with limited dilution method. The growth curve was drawn to analyse its proliferating character in vitro. With Helios gene gun system, DNA vaccine pWRG neu was immunized to 8 week old C57BL/6 mice in the shaved abdominal skin for 3 times at two weekly interval. After immunization, the life span was analyzed. Using MTT assay, the cytolysis activity of the DNA immunized mice spleen cells was compared. Results One clone of neu overexpressed B16F10 neu was selected and its proliferating character was the same as B16F10 and B16F10 pcDNA. In prophylactic, treatment and anti lung metastasis experiments, gene gun mediated pWRG neu immunization could exhibit antitumor effects. The growth and metastasis of neu overexpressed melanoma was reduced dramatically. The spleen cells of the immuned mice showed cytotoxic T lymphocyte (CTL) activity. Conclusion Gene gun mediated gene transfer is effective and practicable. DNA vaccine pWRG neu is potent in preventing subsequent tumor cells challenge, inhibiting the tumor growth and metastasis. [

关 键 词：NEU基因 DNA疫苗 基因枪 抗肿瘤作用 

分 类 号：R730.3[医药卫生—肿瘤]