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利用18型人乳头瘤病毒DNA通过脂质体转染介导对正常及恶性来源的人前列腺上皮细胞进行永生化处理

Lipofection-mediated immortalization of human prostatic epithelial cells of normal and malignant origin using human papillomavirus type 18 DNA.

作者信息

Weijerman P C, König J J, Wong S T, Niesters H G, Peehl D M

机构信息

Department of Urology, Stanford University School of Medicine, California 94305.

出版信息

Cancer Res. 1994 Nov 1;54(21):5579-83.

PMID:7923200
Abstract

Human papillomavirus (HPV) type 18 DNA was introduced into epithelial cell strains derived from normal and cancer tissues of human prostatectomy specimens by the lipofection transfection method. Two cell lines were established: PZ-HPV-7 (transfected normal cell) and CA-HPV-10 (transfected cancer-derived cell). These lines have been maintained for over 100 passages. Incorporation of HPV type 18 DNA was confirmed by polymerase chain reaction. Immunocytochemical analysis showed expression of keratins 5 and 8, similar to the cells of origin, and the early region 6 oncoprotein of HPV. PZ-HPV-7, derived from normal diploid cells, had a modal chromosome number of 46 in early passages but became tetraploid later. CA-HPV-10 cells were aneuploid, and some retained the double minute chromosomes that were noted in the cancer-derived cells of origin. The cell lines showed a typical transformed morphology and were nontumorigenic in nude mice. We conclude that human prostatic epithelial cells derived from both normal and cancer tissues have been successfully transformed to immortality with HPV type 18 DNA. The establishment of these cell lines provides an opportunity for further development of an in vitro model of carcinogenesis for prostate cancer.

摘要

通过脂质体转染法将人乳头瘤病毒18型(HPV-18)DNA导入取自前列腺切除标本正常组织和癌组织的上皮细胞系。建立了两个细胞系:PZ-HPV-7(转染的正常细胞)和CA-HPV-10(转染的癌源细胞)。这些细胞系已传代培养100多代。通过聚合酶链反应确认了HPV-18 DNA的整合。免疫细胞化学分析显示角蛋白5和8的表达,与起源细胞相似,以及HPV早期区域6癌蛋白的表达。源自正常二倍体细胞的PZ-HPV-7在传代早期的众数染色体数为46,但后来变为四倍体。CA-HPV-10细胞为非整倍体,一些细胞保留了起源癌源细胞中观察到的双微小染色体。这些细胞系呈现典型的转化形态,在裸鼠中无致瘤性。我们得出结论,源自正常组织和癌组织的人前列腺上皮细胞已成功地被HPV-18 DNA转化为永生化细胞。这些细胞系的建立为进一步开发前列腺癌体外致癌模型提供了机会。

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