title: Creation of immortalised epithelial cells from ovarian endometrioma.
abstract: BACKGROUND
  Epithelial cells of endometriotic tissues are difficult to propagate in vitro as experimental material is scarce owing to their limited life span. However, there is an increasing concern regarding their malignant transformation in ovaries. The present study sought to generate their stable culture system.
  
  
  RESULTS
  Purified epithelial cells isolated from ovarian endometriomas using microscopic manipulation were successfully immortalised by combinatorial transfection of human cyclinD1, cdk4 and human telomerase reverse transcriptase (hTERT) genes, whereas the introduction of hTERT alone, or together with cdk4, was insufficient for immortalisation, leading to cellular senescence. We confirmed stable cytokeratin expression in the immortalised cells, proving their epithelial origin. These cells expressed progesterone receptor B and showed significant growth inhibition by various progestins. Oestrogen receptor (ER) expression was detected in these cells, albeit at low levels. Additional overexpression of ERα generated stable cells with oestrogen-dependent growth activation. Soft-agar colony formation assay and nude mice xenograft experiments demonstrated that these cells, even those with additional inactivation of p53, did not have transformed phenotypes.
  
  
  CONCLUSIONS
  We for the first time generated immortalised epithelial cells from ovarian endometrioma that retained sex steroid responsiveness. These cells are invaluable tools not only for the consistent in vitro work but also for the study of molecular pathogenesis or carcinogenesis of endometriosis.
authors.affiliation: Department of Obstetrics and Gynecology, Kanazawa University Graduate School of Medical Science, 13-1 Takaramachi, Kanazawa, Ishikawa 920-8641, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Bono
authors.forename: S
authors.initials: S
authors.lastname: Kyo
authors.forename: M
authors.initials: M
authors.lastname: Takakura
authors.forename: Y
authors.initials: Y
authors.lastname: Maida
authors.forename: Y
authors.initials: Y
authors.lastname: Mizumoto
authors.forename: M
authors.initials: M
authors.lastname: Nakamura
authors.forename: K
authors.initials: K
authors.lastname: Nomura
authors.forename: T
authors.initials: T
authors.lastname: Kiyono
authors.forename: M
authors.initials: M
authors.lastname: Inoue
mesh_terms: D000328:Adult; D000818:Animals; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor / Q000378:metabolism / Q000502:physiology* / Q000637:transplantation; D049109:Cell Proliferation; D004715:Endometriosis / Q000201:enzymology / Q000378:metabolism / Q000473:pathology*; D004847:Epithelial Cells / Q000201:enzymology / Q000378:metabolism / Q000473:pathology*; D047628:Estrogen Receptor alpha / Q000235:genetics / Q000378:metabolism; D004967:Estrogens / Q000494:pharmacology / Q000502:physiology; D005260:Female; D015870:Gene Expression; D006801:Humans; D053533:Keratin-8 / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D009368:Neoplasm Transplantation; D015260:Neprilysin / Q000235:genetics / Q000378:metabolism; D010051:Ovarian Neoplasms / Q000201:enzymology / Q000378:metabolism / Q000473:pathology*; D011372:Progestins / Q000494:pharmacology / Q000502:physiology; D011980:Receptors, Progesterone / Q000235:genetics / Q000378:metabolism; D000071999:S100 Calcium-Binding Protein A4
chemical_list: D002135:Calcium-Binding Proteins; C506487:ESR1 protein, human; D047628:Estrogen Receptor alpha; D004967:Estrogens; D053533:Keratin-8; D011372:Progestins; D011980:Receptors, Progesterone; D000071999:S100 Calcium-Binding Protein A4; C097314:progesterone receptor B; C077591:S100A4 protein, human; D015260:Neprilysin