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通过感染SV40或腺病毒12 - SV40杂交病毒,或通过与含有SV40早期区域基因的质粒进行磷酸锶共沉淀转染来实现人支气管上皮细胞的转化。

Transformation of human bronchial epithelial cells by infection with SV40 or adenovirus-12 SV40 hybrid virus, or transfection via strontium phosphate coprecipitation with a plasmid containing SV40 early region genes.

作者信息

Reddel R R, Ke Y, Gerwin B I, McMenamin M G, Lechner J F, Su R T, Brash D E, Park J B, Rhim J S, Harris C C

机构信息

Laboratory of Human Carcinogenesis, National Cancer Institute, Bethesda, Maryland 20892.

出版信息

Cancer Res. 1988 Apr 1;48(7):1904-9.

PMID:2450641
Abstract

Normal human bronchial epithelial cells were infected with SV40 virus or an adenovirus 12-SV40 hybrid virus, or transfected via strontium phosphate coprecipitation with plasmids containing the SV40 early region genes. Colonies of morphologically altered cells were isolated and cultured; these cells had extended culture lifespans compared to normal human bronchial epithelial cells. All cultures eventually underwent senescence, with the exception of one which appears to have unlimited proliferative potential. Colonies arising after viral infection were screened for virus production by cocultivation with Vero cells; only viral nonproducer cultures were analyzed further. The cells retained electron microscopic features of epithelial cells, and keratin and SV40 T-antigen were detected by indirect immunofluorescence. All of the cultures were aneuploid with karyotypic abnormalities characteristic of SV40-transformed cells. No tumors formed after s.c. injection of the cells in nude mice. These cells should be useful for studies of multistage bronchial epithelial carcinogenesis.

摘要

正常人支气管上皮细胞用SV40病毒或腺病毒12 - SV40杂交病毒感染,或通过磷酸锶共沉淀法用含SV40早期区域基因的质粒转染。分离并培养形态改变的细胞集落;与正常人支气管上皮细胞相比,这些细胞具有延长的培养寿命。所有培养物最终都经历衰老,只有一个似乎具有无限增殖潜能的培养物除外。通过与非洲绿猴肾细胞共培养筛选病毒感染后产生的集落中的病毒产生情况;仅对不产生病毒的培养物进行进一步分析。这些细胞保留了上皮细胞的电子显微镜特征,通过间接免疫荧光检测到角蛋白和SV40 T抗原。所有培养物均为非整倍体,具有SV40转化细胞特征性的核型异常。将这些细胞皮下注射到裸鼠中未形成肿瘤。这些细胞对于多阶段支气管上皮细胞癌变的研究应该是有用的。

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