论文部分内容阅读
目的:研究SV40(simianvirus40)病毒对人角朊细胞的体外转化作用以及转化后细胞生物学特性的改变.方法:采用体外共培养法以SV40野生型病毒感染人包皮角朊细胞,观察病毒基因在细胞内的表达以及细胞表型的改变情况.结果:分离获得了可长期体外培养的SV40转化细胞克隆,已传23代以上,体外存活超过250d;DNA印迹实验显示SV40大T基因已整合至细胞染色体内,间接免疫荧光检测示多数细胞胞核内有大T基因产物的表达;转化细胞对裸鼠无致瘤性,并可正常表达角蛋白.结论:SV40病毒转化人角朊细胞后可使其在体外长期培养,该细胞的获得为进一步研究外源性基因对角朊细胞生长的调控奠定了基础.
Objective: To investigate the in vitro transformation of human keratinocyte (SV40) virus by simianvirus40 and the change of cell biological characteristics after transformation. Methods: Human foreskin keratinocytes were infected with SV40 wild-type virus by in vitro co-culture method to observe the expression of the virus gene in the cells and the changes of cell phenotypes. Results: The SV40 transformed cell clone was obtained in vitro for more than 23 passages and survived in vitro for more than 250 days. Southern blotting showed that the SV40 large T gene was integrated into the chromosome of the cell, and indirect immunofluorescence assay showed that most of the cell nuclei There is a large T gene product expression; transformed cells on tumor-free nude mice, and normal expression of keratin. CONCLUSION: SV40 virus can transform human keratinocytes into long-term culture in vitro. The obtained cells provide the basis for further study on the regulation of keratinocyte growth by exogenous genes.