白喉毒素A基因转染对人肺腺癌细胞体内外生长的影响  

Experimental study on human lung adnocarcinoma gene therapy with diphtheria toxin A gene

作　　者：李玉光[1] 张罗献[1] 阎俊宇[2] 马利军[1] 马厚志[1] 

机构地区：[1]河南省人民医院东院呼吸科,郑州450003 [2]郑州大学外语学院,郑州450052

出　　处：《郑州大学学报（医学版）》2006年第3期535-537,共3页Journal of Zhengzhou University(Medical Sciences)

摘　　要：目的:探讨白喉毒素A(DTA)基因在肺腺癌基因治疗中的作用。方法:①体外细胞毒试验:对转染DTA基因的人肺腺癌细胞A549(A549/DTA)、转染空载体的人肺腺癌细胞A549(A549/DTA-)和野生型A549,采用MTT法测定四环素作用72h后的细胞存活率。②动物实验:将1.2×107的A549细胞接种于裸鼠皮下,当肿瘤长至约350mg时,随机分组:转DTA基因治疗组,荷瘤裸鼠每次给予含DTA基因的病毒上清0.4ml瘤体注射;空载体组给予0.4ml含空载体的病毒上清;对照组给予0.4ml生理盐水,均2d1次,连续3周;从开始注射病毒上清之日起,每周2次测量肿瘤大小,按公式计算肿瘤相对质量。3周后处死动物,瘤组织作病理分析。结果:①细胞存活率:转空载体的A549与野生型A549形态无明显改变,而转染DTA基因的A549细胞体积明显缩小,并有细胞碎片。②瘤体注射DTA基因的病毒上清可使皮下肿瘤显著缩小,从注射之日起至21d,肿瘤体积缩小,而空载体组、对照组在相同时间内肿瘤体积增加。转DTA基因治疗的瘤组织出现了细胞坏死和凋亡,而空载体组及对照组瘤组织无坏死及细胞凋亡征象。结论:四环素调控白喉毒素A基因对肺腺癌有一定的治疗作用。Aim ： To investigate the effect of gene therapy with diphtheria toxin A gene （DTA） on human lung adenocarcinoma A549 cells. Methods： The A549 cells transfected with DTA （A549/DTA） , A549 cells transfected with mock vector （A549/DTA^- ） and wild type A549 cells were treated with tetracycline for 72.h, and the cell survival rate was determined with MTT; the nude mice bearing A549 cell were divided into 3 groups when the tumor was about 350 mg. Group Ⅰ , the animals were injected in tumor with viral solution of A549/DTA for three weeks. Group Ⅱ ,the animals were injected equal volume of viral solution of A549/DTA^-. Group Ⅲ ,the animals were injected equal volume of normal saline. In this period,the tumor weight was measured and the size of tumor was calculated twice a week. The animals were killed and tumor samples were prepared for pathology observation. Results： The survival rate of A549/DTA cells was apparently lower than those of A549 cells and the A549/DTA - cells. The tumor size dcreased obviously in group Ⅰ , while the tumors in group Ⅱ and Ⅲ increased 3 times during the three weeks. Cell necrosis and apoptosis appeared in tumor tissue of group Ⅰ ,while there was no necrosis and apoptosis in group Ⅱ and Ⅲ . Conclusion： DTA gene regulated by tetracycline has therapeutic effects on lung adenocarcinoma.

关 键 词：人肺腺癌细胞 白喉毒素A基因 基因治疗 

分 类 号：R734.2[医药卫生—肿瘤]