Molecular and cytogenetic analysis of immortalized human primary keratinocytes obtained after transfection with human papillomavirus type 16 DNA.
Molecular and cytogenetic analysis of immortalized human primary keratinocytes obtained after transfection with human papillomavirus type 16 DNA.
复制标题
对转染人乳头瘤病毒 16 型 DNA 后获得的永生化人原代角质形成细胞进行分子和细胞遗传学分析。
作者：
M. Durst;R. T. Dzarlieva;P. Boukamp;N. Fusenig;L. Gissmann
人包皮角质形成细胞在人乳头瘤病毒(HPV)16型DNA转染后获得了增殖的群体(目前处于第六十代)。相比之下，对照培养物没有存活到第六代以后。第12代细胞的细胞遗传学分析显示为异倍体雄性核型。在大约50%的细胞中发现了共同的标记染色体，这表明至少有一部分群体是克隆起源的。对细胞DNA的Southern印迹分析进一步证实了这一点，该分析显示，低聚HPV16基因组整合在宿主DNA中的单一位置。从HPV16基因组早期区域转录的RNA存在于细胞质中。HPV16DNA对人角质形成细胞的永生作用可以在第二次实验中重现。这些细胞系代表了一种独特的系统，可以在体外研究HPV与其自然靶细胞的相互作用。
A proliferating population of human foreskin keratinocytes (presently in the sixtieth passage) has been obtained after transfection with human papillomavirus (HPV) type 16 DNA. In contrast, the control cultures did not survive beyond the sixth passage. Cytogenetic analysis of cells taken from the twelfth passage revealed a heteroploid male karyotype. In approximately 50% of the cells a common marker chromosome was found, suggesting a clonal origin for at least part of the population. This is further substantiated by Southern blot analysis of cellular DNA which revealed oligomeric HPV 16 genomes integrated at a single site within the host DNA. RNA transcribed from the early region of the HPV 16 genome was identified in the cytoplasm. The immortalizing effect of HPV 16 DNA on human keratinocytes could be reproduced in a second experiment. Such cell lines represent an unique system to study the interaction of HPV with its natural target cell in vitro.