Oncogenic transformation of human ovarian surface epithelial cells with defined cellular oncogenes

Sasaki, Rumi; Narisawa‐Saito, Mako; Yugawa, Takashi; Fujita, Masatoshi; Tashiro, Hironori; Katabuchi, Hidetaka; Kiyono, Tohru

doi:10.1093/carcin/bgp007

Cited by 96 publications

(93 citation statements)

References 45 publications

(54 reference statements)

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“…Few models of human ovarian cancer can produce high-grade serous carcinomas using clinically relevant genetic alterations. Although two groups have reported high-grade serous tumor formation with human OSE-based models (33)(34)(35), their studies either did not demonstrate a Mülle-rian phenotype or relied on SV40 TAg expression to achieve transformation.…”

Section: R24cmentioning

confidence: 99%

Modeling high-grade serous ovarian carcinogenesis from the fallopian tube

Karst

Levanon

Drapkin

2011

Proc. Natl. Acad. Sci. U.S.A.

255

281

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High-grade serous ovarian carcinoma (HGSOC) is a lethal disease for which improved screening and treatment strategies are urgently needed. Progress in these areas is impeded by our poor understanding of HGSOC pathogenesis. Most ovarian cancer research is based on the hypothesis that HGSOC arises from ovarian surface epithelial cells. However, recent studies suggest that >50% of high-grade serous carcinomas involving the ovary likely arise from fallopian tube epithelium. Therefore, limiting HGSOC research to modeling based on ovarian surface epithelium alone is inadequate. To address the need for a fallopian tube-based model of HGSOC, we have developed a system for studying human fallopian tube secretory epithelial cell (FTSEC) transformation. Our model is based on (i) immortalization of FTSECs isolated from primary samples of normal, nondiseased human fallopian tubes, (ii) transformation of FTSECs with defined genetic elements, and (iii) xenograft-based tumorigenic assays. We use our model to show that FTSECs immortalized with human telomerase reverse transcriptase (hTERT) plus SV40 large T and small T antigens are transformed by either oncogenic Ras (H-Ras V12 ) or c-Myc expression, leading to increased proliferation, clonogenicity, and anchorage-independent growth. Additionally, we demonstrate that FTSECs remain susceptible to c-Myc-mediated transformation in the absence of viral oncoproteins, by replacing SV40 large T and small T antigens with sh-p53, mutant CDK4 (CDK4 R24C ), and sh-PP2A-B56γ. Importantly, all transformed FTSECs gave rise to highgrade Müllerian carcinomas that were grossly, histologically, immunophenotypically, and genomically similar to human HGSOC. With this model, we will now be able to assess the transformative effects of specific genetic alterations on FTSECs in order to characterize their respective roles in HGSOC development.varian cancer is the fifth deadliest cancer among American women (1). It has a disproportionately high mortality rate, attributed primarily to difficulties in diagnosing early stage disease and to the development of drug resistance in tumors that were initially chemosensitive (2). To improve ovarian cancer screening and treatment strategies, we must better understand the cancer's origin and pathogenesis. The most common histologic subtype, accounting for >50% of ovarian epithelial malignancies, is serous ovarian carcinoma (3-5). Because of inadequate early detection tools, the vast majority of serous ovarian carcinomas (>80%) are diagnosed at late stage [International Federation of Gynecology and Obstetrics (FIGO) stages III-IV], for which the 5-y survival rate is only 9-34% (6). Most of these (>50%) are classified as "high-grade" based on their degree of nuclear atypia and high mitotic index (7). High-grade serous ovarian carcinoma (HGSOC) stands out from other subtypes both for its aggressive nature and because it harbors unique genetic alterations. For example, clear cell, endometrioid, low-grade serous, and mucinous ovarian carcinomas typically present as indo...

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Section: R24cmentioning

confidence: 99%

Modeling high-grade serous ovarian carcinogenesis from the fallopian tube

Karst

Levanon

Drapkin

2011

Proc. Natl. Acad. Sci. U.S.A.

255

281

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“…Construction of CSII-CMV-cyclin D1, -CDK4R24C and -hTERT was described previously. 35 The recombinant lentiviruses with the vesicular stomatitis virus G glycoprotein were produced as described previously. 36 The recombinant retroviruses encoding hTERT and E7 were produced as described previously.…”

Section: Viral Vector Construction and Viral Transductionmentioning

confidence: 99%

“…9,37 Hu5, HM1-8 and HM2-5 cells were transduced with recombinant lentiviruses and retroviruses as described. 9,11,35 Following inoculation with viruses, the continuously proliferating cells were selected without drug treatment.…”

Section: Viral Vector Construction and Viral Transductionmentioning

confidence: 99%

CDK4 and cyclin D1 allow human myogenic cells to recapture growth property without compromising differentiation potential

Kiyono²,

et al. 2011

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In vitro culture systems of human myogenic cells contribute greatly to elucidation of the molecular mechanisms underlying terminal myogenic differentiation and symptoms of neuromuscular diseases. However, human myogenic cells have limited ability to proliferate in culture. We have established an improved immortalization protocol for human myogenic cells derived from healthy and diseased muscles; constitutive expression of mutated cyclin-dependent kinase 4, cyclin D1 and telomerase immortalized human myogenic cells. Normal diploid chromosomes were preserved after immortalization. The immortalized human myogenic cells divided as rapidly as primary human myogenic cells during the early passages, and underwent myogenic, osteogenic and adipogenic differentiation under appropriate culture conditions. The immortalized cells contributed to muscle differentiation upon xenotransplantation to immunodeficient mice under conditions of regeneration following muscle injury. We also succeeded in immortalizing cryopreserved human myogenic cells derived from Leigh disease patients following primary culture. Forced expression of the three genes shortened their cell cycle to o30 h, which is similar to the doubling time of primary cultured human myogenic cells during early passages. The immortalization protocol described here allowed human myogenic cells to recapture high proliferation activity without compromising their differentiation potential and normal diploidy.

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“…Thus, while telomerase activity is indispensable in the immortalization step of human OSE, p53 and RB1 are not, at least under certain conditions. Furthermore, TERT-immortalized human OSE cells have lengthened telomeres, are diploid without chromosomal instability, and maintain epithelial characters without tumorigenicity (Kusakari et al 2003, Yang et al 2007a, 2007b, Sasaki et al 2009). …”

Section: Reactivation Of Telomerase In Ose and The Role In Ovarian Rementioning

confidence: 99%

“…Consistently, TERT-and SV40 large T antigen-immortalized human OSE cells undergo anchorage-independent growth in vitro and tumorigenesis in mice upon forced expression of either oncogene ERBB2 (c-erbB-2) or mutant HRAS (Ha-Ras; Kusakari et al 2003). Recently, oncogenic transformation of human OSE cells has been investigated with defined cellular oncogenes (Sasaki et al 2009). Non-viral transduction of primary human OSE cells with human genes encoding mutant cyclindependent kinase 4, cyclinD1, and TERT establishes normal diploid OSE cells with extended proliferation without chromosomal instability (Sasaki et al 2009).…”

Section: Reactivation Of Telomerase In Ose and The Role In Ovarian Rementioning

confidence: 99%

Telomerase in the ovary

Liu¹,

Li²

2010

Reproduction

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Telomerase, an enzyme complex that binds the chromosome ends (telomeres) and maintains telomere length and integrity, is present in germ cells, proliferative granulosa cells, germline stem cells, and neoplastic cells in the ovary, but it is absent in differentiated or aged cells. Activation of telomerase in the ovary underpins both benign and malignant cell proliferation in several compartments, including the germ cells, membrana granulosa, and the ovarian surface epithelium. The difference in telomerase operation between normal and abnormal cell proliferations may lie in the mechanisms of telomerase activation in a deregulated manner. Recent studies have implicated telomerase activity in ovarian cancer as well as oogenesis and fertility. Inhibition of telomerase and the shortening of telomeres are seen in occult ovarian insufficiency. Studies of how telomerase operates and regulates ovary development may provide insight into the development of both germ cells for ovarian reproductive function and neoplastic cells in ovarian cancer. The current review summarizes the roles of telomerase in the development of oocytes and proliferation of granulosa cells during folliculogenesis and in the process of tumorigenesis. It also describes the regulation of telomerase by estrogen in the ovary.

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Oncogenic transformation of human ovarian surface epithelial cells with defined cellular oncogenes

Cited by 96 publications

References 45 publications

Modeling high-grade serous ovarian carcinogenesis from the fallopian tube

Modeling high-grade serous ovarian carcinogenesis from the fallopian tube

CDK4 and cyclin D1 allow human myogenic cells to recapture growth property without compromising differentiation potential

Telomerase in the ovary

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