CHARACTERIZATION OF HUMAN OVARIAN SURFACE EPITHELIAL-CELLS IMMORTALIZED BY HUMAN PAPILLOMA VIRAL ONCOGENES (HPV-E6E7 ORFS)
CHARACTERIZATION OF HUMAN OVARIAN SURFACE EPITHELIAL-CELLS IMMORTALIZED BY HUMAN PAPILLOMA VIRAL ONCOGENES (HPV-E6E7 ORFS)
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DOI：
10.1006/excr.1995.1184
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发表时间：
1995-06-01
影响因子：
3.7
通讯作者：
BERKOWITZ, RS
中科院分区：
文献类型：
--
作者：
TSAO, SW;MOK, SC;BERKOWITZ, RS
通过表达HPV-E6 E7开放阅读框（ORF）的逆转录病毒载体（LXSN-16 E6 E7）使原代人卵巢表面上皮（HOSE）细胞永生化。三次尝试中有三次实现了原代卵巢上皮细胞的永生化。在一个细胞系HOSE 6-3中进行详细分析，该细胞系是根据其上皮形态学选择的。在传代培养20次时检查时，永生化细胞系（HOSE 6-3）在裸鼠中无致瘤性。细胞遗传学分析证实了它的人类起源，详细的核型分析显示，由约60%的二倍体和40%的近四倍体细胞组成的混合核型。免疫荧光和双向凝胶电泳显示，在一个细胞亚群中观察到9号环状染色体的克隆性染色体畸变，波形蛋白和几种细胞角蛋白（K7，K8，K18，K19）的存在。HOSE 6-3细胞的细胞骨架丝的轮廓与永生化前的正常卵巢上皮细胞的基本相同。与正常卵巢上皮细胞相比，永生化卵巢上皮细胞对TGF-β 1抑制的敏感性较低。永生化系HOSE 6-3具有改变的生长特性，包括更高的增殖速率、平板接种效率和饱和密度。人卵巢上皮细胞系的建立为研究卵巢癌的发生提供了一个体外模型。(C)出版社：Academic Press
Primary human ovarian surface epithelial (HOSE) cells were immortalized by a retroviral vector (LXSN-16E6E7) expressing HPV-E6E7 open reading frames (ORF). Immortalizations of primary ovarian epithelial cells were achieved in three of three attempts. Detailed analysis was carried out in one line, HOSE 6-3, selected on the basis of its epithelial morphology. The immortalized line (HOSE 6-3) was nontumorigenic in nude mice when examined at subculture number 20. Cytogenetic analysis confirmed its human origin and detailed karyotypic analysis revealed a mixed karyotype made up of about 60% of diploid and 40% of near-tetraploid cells. Clonal chromosomal aberration was observed in a subpopulation of cells involving a ring chromosome number 9, Immunofluorescence and two-dimensional gel electrophoresis revealed the presence of vimentin and several species of cytokeratin (K7, K8, K18, K19). The profile of the cytoskeletal filaments of HOSE 6-3 cells is largely identical with that of normal ovarian epithelial cells before immortalization. The immortalized ovarian epithelial cells have a lower sensitivity to TGF-beta 1 inhibition compared to normal ovarian epithelial cells. The immortalized line, HOSE 6-3, has altered growth properties including a higher proliferation rate, plating efficiency, and saturation density. The establishment of a continuous line of human ovarian epithelial cells may provide an in vitro model for study of carcinogenesis in human ovarian cancers. (C) 1995 Academic Press, Inc.