Immortalization of human urothelial cells by human papillomavirus type 16 E6 and E7 genes in a defined serum-free system.
Immortalization of human urothelial cells by human papillomavirus type 16 E6 and E7 genes in a defined serum-free system.
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在确定的无血清系统中，人乳头瘤病毒 16 型 E6 和 E7 基因使人尿路上皮细胞永生化。
DOI：
10.1111/j.1365-2184.2007.00428.x
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发表时间：
2007
影响因子：
8.5
通讯作者：
Bassuk,JA
中科院分区：
文献类型：
--
作者：
Carmean,N;Kosman,JW;Leaf,EM;Hudson,AE;Opheim,KE;Bassuk,JA
正常的人上皮细胞培养在体外表现出有限的寿命(尽管不同组织之间不同)。在以前的研究中，尿路上皮细胞的永生化是通过逆转录病毒转化人乳头瘤病毒16型E6和E7基因，在含有血清或牛垂体提取物的未确定的培养体系中进行的。目的：由于此类培养体系的结果的多样性，我们开发了一种使用定义的无血清培养系统来永生化尿路上皮细胞的方法。方法和结果：通过逆转录病毒转化人乳头瘤病毒16型E6和E7永生化是成功的，转化的尿路上皮细胞可以延长正常寿命并恢复端粒酶活性。转化细胞保持了典型的形态，并表现出与未转化细胞相似的生长速度、细胞角蛋白免疫反应模式和对生长因子的反应。核型分析显示，在人类乳头瘤病毒16型E6和E7型转化的上皮细胞中，与先前报道的突变一致的基因突变逐渐积累。结论：延长细胞体外寿命的能力有可能减少对组织样本的持续需求，并使对一个细胞系的完整研究成为可能。
Normal human epithelial cell cultures exhibit a limited (although different between tissues) lifespanin vitro. In previous studies, urothelial cell cultures were immortalized using retroviral transformation with human papillomavirus type 16 E6 and E7 genes, in undefined culture systems containing serum or bovine pituitary extract.Objective: Due to the variability of results in such systems, we instead developed a procedure for the immortalization of urothelial cells using a defined, serum‐free culture system.Method and results: Immortalization through retroviral transformation with human papillomavirus type 16 E6 and E7 was successful, and transformation of urothelial cells conferred an extended over normal lifespan and restored telomerase activity. Transformed cells retained typical morphology and exhibited a similar growth rate, cytokeratin immunoreactivity pattern, and response to growth factors as observed in untransformed cells. Karyotype analysis revealed a gradual accumulation of genetic mutations that are consistent with previously reported mutations in epithelial cells transformed with human papillomavirus type 16 E6 and E7.Conclusion: The ability to extend thein vitrolifespan of cells holds the potential to reduce the continuous need for tissue samples and to enable complete investigations with one cell line.