转染白喉毒素基因对人肺腺癌细胞的影响

Effect of transfection of diphtheria toxin gene on human lung adenocarcinoma cell

目的 探讨白喉毒素A基因对人肺腺癌细胞 (A5 4 9)的影响。方法 对转染白喉毒素A基因的人肺腺癌细胞 (A5 4 9/DTA) ,转空载体的人肺腺癌细胞 (A5 4 9/DTA-)野生型A5 4 9细胞 ,采用MTT法测定四环素作用 72h后的细胞存活率。动物实验 :12只裸鼠随机分为转基因组、空载体组和对照组 ,每组 4只 ,将 1.2× 10 7A5 4 9/DTA细胞 ,A5 4 9/DTA-细胞及野生型A5 4 9细胞分别接种于相应的裸鼠皮下 ,给裸鼠饮含四环素(1μg/ml)的水 ,成瘤后 ,每周 2次测量瘤体大小 ,计算瘤体重量。 结果 A5 4 9/DTA细胞存活率较A5 4 9/DTA-细胞 ,A5 4 9细胞存活率明显下降 (P <0 .0 1) ,而A5 4 9/DTA-细胞与A5 4 9细胞存活率无差别 (P >0 .0 5 )。空载体组与对照组的裸鼠全部长出肿瘤 ,且肿瘤生长迅速 ,而转基因组有 1只裸鼠未长出肿瘤 ,肿瘤生长缓慢 ,前者瘤体的重量是后者的 3倍。结论 体外转染白喉毒素A基因可杀伤人肺腺癌细胞并降低人肺腺癌细胞的致瘤性。

Objective:To invesigate effect on human lung adenocarcinoma cell by transfection of DTA gene.Methods:The survival rate of A549/DTA(DTA transferred A549) cells, A549/DTA-(empity vector transferred A549) cells and wide type A549 cells were measured with MTT assay after treating all cells by tetracycline for 72 hours .Twelve nude mice were divided into three groups at random,transferring gene group,empity vector group and contrast group,each group has four animals. 1.2×10 7 cells of A549/DTA,A549/DTA - and A549 were inoculated into animal of each group respectively. Animals drank water of tetracycline(1?μg/ml).The tumor weights were measured and calculated with the experience formula twice every week when the animal grew up tumor. Results:The survival rate of A549/DTA cells were dropped significantly than that of A549/DTA-cells and A549cells (P<0.01),but there was no difference insurvival rate between A549/DTA-cells and A549 cells(P>0.05).Nude mice of empity vector group and contrast group were all oncogenesis and tumors grew fast,but there was a nude mouse of transferring gene group without oncogenesis and tumors grew slowly, the average tumor weight of the former is three times as that of the latter's.Conclusion:Transfection of DTA gene in vitro may kill human lung adenocarcinoma cells and reduce the oncogenicity of human lung adenocarcinoma cells.