人肺癌相关抗原基因ALT04-AG表达抑制对人肺癌细胞株生长及基因表达谱的影响

目的探讨抑制人肺癌相关抗原基因ALT04-AG表达对细胞生长特性及相关基因表达的影响。方法重组反义ALT04-AGRNA真核表达质粒,经脂质体介导转染人肺癌细胞株(L78),以MTT、FCM法分析转染细胞生长特性,以Northern b lot、免疫组化染色及基因芯片技术检测相关基因的表达。结果构建了重组表达反义ALT04-AGRNA的真核表达质粒[pALT04-AG(as)],经该质粒转染及二氟甲基鸟氨酸(d ifluorom ethylorn ith ine,DFMO)处理的L78细胞均引起ALT04-AG表达下调及细胞的增殖抑制,后者还导致细胞凋亡比例增加。结论pALT04-AG(as)转染L78细胞或用DFMO抑制多胺生物合成,可通过对相关基因表达的调控促使L78细胞恶性表型逆转,而后者的作用更为广泛。本结果为探索肿瘤的诊断与治疗提供了有意义的线索。

Inhibition of human lung carcinoma-associated antigen gene(ALT04-AG) expression on human lung carcinoma cell line growth and gene expression profile

Objective To study the effect of inhibition of human lung carcinoma-associated antigen gene(ALT04-AG) expression on the growth characteristic and associated gene expression of human lung carcinoma cells(L78).Methods L78 cells were transfected with recombinant plasmid vector expressing(ALT04-AG) antisense RNA mediated by liposome reagent,and treated with DFMO respectively. The growth characteristics were studied by MTT,FCM analysis.The associated gene expression was detected by immunocytochemical staining,Northern blot and gene chip analysis.Results The recombinant plasmid expressing ALT04-AG antisense RNA pALT04-AG as constructed.It was shown that the down-regulation of ALT04-AG gene expression was observed not only in pALT04-AG as transfected L78 cells,but also in DFMO-treated L78 cells,which resulted in growth inhibition of these cells.Gene chip analysis indicated that inhibition of(ALT04AG) gene expression by ALT04-AG antisense RNA regulated some proliferation associated gene expression,while inhibition of polyamine biosynthesis by DFMO regulated proliferation associated genes and also some apoptosis associated genes.Conclusion It is suggested that human lung carcinoma associated antigen gene ALT04-AG plays an essential role in maintaining malignant phenotype of cancer cells.