Characterization and Tumorigenicity of Human Ovarian Surface Epithelial Cells Immortalized by SV40 Large T Antigen

Nitta, Makoto; Katabuchi, Hidetaka; Ohtake, Hideyuki; Tashiro, Hironori; Yamaizumi, Masaru; Okamura, Hitoshi

doi:10.1006/gyno.2000.6084

Cited by 69 publications

(50 citation statements)

References 20 publications

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“…Normal ovarian surface epithelial cell lines (OSE2 and OSE4) were established by one of the authors (M. N.; Ref. 23). Cell lines were maintained in DMEM/F12 (Invitrogen) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (Invitrogen), and were incubated in 5% CO 2 at 37°C.…”

Section: Methodsmentioning

confidence: 99%

Decreased Expression of 14-3-3σ Is Associated with Advanced Disease in Human Epithelial Ovarian Cancer

Akahira

Sugihashi²,

Suzuki

et al. 2004

Clinical Cancer Research

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101

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Purpose: In this study, we examined the promoter methylation status and expression of 14-3-3 and evaluated its clinical significance in epithelial ovarian cancer.Experimental Design: Twelve ovarian cancer cell lines; 2 ovarian surface epithelial cell lines; and 8 normal, 8 benign, 12 borderline, and 102 ovarian cancer tissues were examined. Methylation-specific PCR, quantitative reverse transcription-PCR, and immunohistochemistry were used to evaluate methylation status and expression of 14-3-3 gene and protein.Results: Among the 12 ovarian cancer cell lines, the presence of a methylated band was detected in seven cell lines. Median values of relative 14-3-3 gene expression in cancers with methylation (3.27) were significantly lower than those without methylation (16.4; P < 0.001). Treatment of 5-aza-2-deoxycitidine resulted in the demethylation of the promoter CpG islands and reexpression. All of the normal, benign, and borderline tissues were positive for 14-3-3 protein, and in ovarian cancer tissues, 73.5% (75 of 102) were positive for 14-3-3 protein and was almost consistent with methylation status. Negative immunoreactivity of 14-3-3 was significantly correlated with high age and serous histology, high-grade, advanced-stage residual tumor of >2 cm, high serum CA125, high Ki-67 labeling index, and positive p53 immunoreactivity. 14-3-3 immunoreactivity was significantly associated with overall survival (P ‫؍‬ 0.0058).Conclusions: Our findings suggest that 14-3-3 is inactivated mainly by aberrant DNA methylation and that it may play an important role in the pathogenesis of epithelial ovarian cancer.

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Section: Methodsmentioning

confidence: 99%

Decreased Expression of 14-3-3σ Is Associated with Advanced Disease in Human Epithelial Ovarian Cancer

Akahira

Sugihashi²,

Suzuki

et al. 2004

Clinical Cancer Research

Self Cite

101

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“…In addition, adequate experimental systems and suitable animal models are lacking due to spontaneous epithelial ovarian tumours being extremely rare in nonprimate mammals, including mice. During the last decade, the isolation, culture, and immortalisation of human OSE cells was developed (Maines-Bandiera et al, 1992;Tsao et al, 1995;Nitta et al, 2001). Using the experimental system of Auersperg et al (1999), E-cadherin expression was recently addressed in human ovarian carcinogenesis (Ong et al, 2000).…”

mentioning

confidence: 99%

“…Until now, using human OSE cells, two kinds of immortalised cell lines were established by introduction of SV40 large T antigen (LT) (Maines-Bandiera et al, 1992;Nitta et al, 2001) or by human papillomavirus (HPV) E6 and E7 (E6/E7) transduction (Tsao et al, 1995). The OSE cells transduced by SV40 LT and by E6/E7 had chromosomal alterations ranging in number from 50 to 69 (Nitta et al, 2001) and 43 to 88 (Tsao et al, 1995), respectively. These OSE cells were not thought to have active telomerase, and consequently resulted in chromosomal alterations.…”

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confidence: 99%

Establishment of an immortalised human ovarian surface epithelial cell line without chromosomal instability

et al. 2005

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Epithelial ovarian carcinoma is thought to derive from ovarian surface epithelium (OSE). The black box of the early molecular changes in ovarian carcinogenesis is being interpreted by the development of experimental systems employing immortalised human OSE cells. However, the existing cell lines of the OSE cells have limited utility due to chromosomal instability. Our goal was to establish new immortalised human OSE cells that retain the original characteristics of the primary cells without chromosomal alterations. Using primary human OSE cells obtained from a postmenopausal patient with endometrial cancer, five cell lines ('HOSE1' lines) were newly established by infection with retroviral expression vectors containing type 16 human papillomavirus (HPV-16) E6, E7, a variant E6 (E6D151), and Bmi1 polycomb gene, in combination with telomerase reverse transcriptase (hTERT). Consequently, five HOSE1s cell lines, HOSE1s-E6/hTERT, -E7/hTERT, -E6/E7/hTERT, -E6D151/E7/hTERT, and -E6D151/Bmi1/hTERT, grew beyond the population doubling number of 200. These cell lines, except for HOSE1-E6/hTERT, essentially showed the original features of the primary human OSE cells. Of them, HOSE1-E7/hTERT preserved diploidy in a kariotype analysis, and did not show transformed phenotypes in anchorage-independent growth and tumour formation. Thus, HOSE1-E7/hTERT may provide a novel model system with which to investigate the mechanisms of early molecular changes.

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“…Human OSE cells provide a useful experimental system for the analysis of ovarian carcinogenesis. Two studies on immortalization of human OSE cells by the introduction of LT alone have been already reported (MainesBandiera et al, 1992;Nitta et al, 2001). Maines-Bandiera et al reported that OSE lines transfected with LT exhibited an extended life span, but subsequently underwent a progressive reduction of growth and characteristic morphologic changes associated with senescence within 20 passages.…”

Section: Discussionmentioning

confidence: 99%

“…Two studies on malignant transformation of human OSE cells by gene transfection have been reported (Ong et al, 2000;Nitta et al, 2001). A recent report demonstrated that transfection of human telomerase reverse transcriptase (hTERT) into SV40 large T antigen (LT)-transfected human somatic cells showed 100% immortalization in transfected cells (Hahn et al, 1999).…”

mentioning

confidence: 99%

C-erbB-2 or mutant Ha-ras induced malignant transformation of immortalized human ovarian surface epithelial cells in vitro

et al. 2003

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Ovarian cancer is believed to develop from the ovarian surface epithelium through the accumulation of aberrations of oncogenes and/or tumor suppressor genes. However, it is unclear how the gene abnormalities are involved in ovarian carcinogenesis. To elucidate the process, we transfected genes reported to show their abnormalities in human ovarian cancers into human ovarian surface epithelial cells. Immortalization of the cells was achieved by the transfection of SV40 large T antigen (LT) and human telomerase reverse transcriptase (hTERT); however, the resultant cells showed no tumorigenesis. Additional transfection of either cerbB-2 or mutant Ha-ras into the immortalized cells showed the anchorage-independent growth and tumorigenesis in mice with the incidence of 50% and 40%, respectively. Histologically, all the tumours were undifferentiated. In association with the tumorigenesis, the cells expressing c-erbB-2 or mutant Ha-ras demonstrated increased vascular endothelial growth factor secretion under hypoxia and enhanced resistance to apoptosis compared with the immortalized cells. Collectively, the introduction of either c-erbB-2 or mutant Ha-ras in the cells, which were efficiently immortalized by the transfection of LT and hTERT, showed tumorigenicity, suggesting that c-erbB-2 or mutant Ha-ras genes might be involved in ovarian carcinogenesis.

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Characterization and Tumorigenicity of Human Ovarian Surface Epithelial Cells Immortalized by SV40 Large T Antigen

Cited by 69 publications

References 20 publications

Decreased Expression of 14-3-3σ Is Associated with Advanced Disease in Human Epithelial Ovarian Cancer

Decreased Expression of 14-3-3σ Is Associated with Advanced Disease in Human Epithelial Ovarian Cancer

Establishment of an immortalised human ovarian surface epithelial cell line without chromosomal instability

C-erbB-2 or mutant Ha-ras induced malignant transformation of immortalized human ovarian surface epithelial cells in vitro

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