Abstract
Human epithelial ovarian neoplasm is well‐known to be sex steroid‐related, but the possible biological significance of progesterone actions in these tumors remains controversial. In this study, we examined the differential expression patterns of the two progesterone receptor (PR) isoforms, PRA and PRB, using immunohistochemistry and real‐tune quantitative RT‐PCR in normal and neoplastic ovarian tissues, and in cell lines derived from a normal ovarian surface epithelium and an ovarian epithelial carcinoma in order to further elucidate the possible involvement of progesterone in the development of ovarian neoplasms. The median H scores for PR isoforms in normal (n=8), benign (n=10), borderline (n=8) and malignant (n=24) ovarian tissues were as follows; PRA: 194.0, 171.0, 49.5, 0 (P<0.05), and PRB: 175.0, 180.5, 251.5, 168.5, respectively. In ovarian cancer cell lines (OVCAR–3 and Caov–3), the PRB/PRAB mRNA ratio was increased by 17β‐estradiol, both tune‐and dose‐dependently. However, this ratio was unaltered following the addition of 17β‐estradiol in a normal ovarian epithelial cell line (NOV–31). Immunoblotting analysis demonstrated that PRB protein expression was markedly up‐regulated in OVCAR–3, whereas the PRA and PRB isoforms both appeared to be increased in NOV–31. These results suggest that down‐regulation of PRA is associated with the development of ovarian epithelial carcinoma.
Keywords: Ovary, Cancer, Immunohistochemistry, Quantitative RT‐PCR, Progesterone receptor
Full Text
The Full Text of this article is available as a PDF (3.6 MB).
REFERENCES
- 1. ) Akahira , J. , Yoshikawa , H. , Shimizu , Y. , Tsunematsu , R. , Hirakawa , T. , Kuramoto , H. , Shiromizu , K. , Kuzuya , K. , Kamura , T. , Kikuchi , Y. , Kodama , S. , Yamamoto , K. and Sato , S.Prognostic factors of stage IV epithelial ovarian cancer: a multicenter retrospective study . Gynecol. Oncol. , 81 , 398 – 403 ( 2001. ). [DOI] [PubMed] [Google Scholar]
- 2. ) Boring , C. C. , Squires , T. S. , Tong , T. and Montgomery , S.Cancer statistics . CA Cancer J. Clin. , 44 , 7 – 26 ( 1994. ). [DOI] [PubMed] [Google Scholar]
- 3. ) Rao , B. R. and Slotman , B. J.Endocrine factors in common epithelial ovarian cancer . Endocr. Rev. , 12 , 14 – 26 ( 1991. ). [DOI] [PubMed] [Google Scholar]
- 4. ) Lantta , M.Estradiol and progesterone receptors in normal ovary and ovarian tumors . Acta Obstet. Gynecol. Scand. , 63 , 497 – 503 ( 1984. ). [DOI] [PubMed] [Google Scholar]
- 5. ) Willcocks , D. , Toppila , M. , Hudson , C. N. , Tyler , J. P. P. , Baird , P. J. and Eastman , C. J.Estrogen and progesterone receptors in human ovarian tumors . Gynecol. Oncol. , 16 , 246 – 253 ( 1983. ). [DOI] [PubMed] [Google Scholar]
- 6. ) Risch , H. A.Hormonal etiology of epithelial ovarian cancer, with a hypothesis concerning the role of androgens and progesterone . J. Natl. Cancer Inst. , 90 , 1774 – 1786 ( 1998. ). [DOI] [PubMed] [Google Scholar]
- 7. ) Garg , P. P. , Kerlikowske , K. , Subak , L. and Grady , D.Hormone replacement therapy and the risk of epithelial ovarian carcinoma: a meta‐analysis . Obstet. Gynecol. , 92 , 472 – 479 ( 1998. ). [DOI] [PubMed] [Google Scholar]
- 8. ) Schneider , H. P. and Birkhauser , M.Does HRT modify risk of gynecological cancers ? Int. J. Fertil. Menopausal Stud. , 40 , 40 – 53 ( 1998. ). [PubMed] [Google Scholar]
- 9. ) Mahlck , C. G. , Backstrom , T. and Kjellgren , O.Plasma level of estradiol in patients with ovarian malignant tumors . Gynecol. Oncol. , 30 , 313 – 320 ( 1988. ). [DOI] [PubMed] [Google Scholar]
- 10. ) Modan , B. , Ron , E. , Lerner‐Geva , L. , Blumstein , T. , Menczer , J. , Rabinovici , J. , Oelsner , G. , Freedman , L. , Mashiach , S. and Lunenfeld , B.Cancer incidence in a cohort of infertile women . Am. J. Epidemiol. , 147 , 1038 – 1042 ( 1998. ). [DOI] [PubMed] [Google Scholar]
- 11. ) Evans , R. M.The steroid and thyroid hormone receptor superfamily . Science , 240 , 889 – 895 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 12. ) Horwitz , K. B. and Alexander , P. S.In situ photolinked nuclear progesterone receptors of human breast cancer cell lines: subunit molecular weights after transformation and translocation . Endocrinology , 113 , 2195 – 2201 ( 1983. ). [DOI] [PubMed] [Google Scholar]
- 13. ) Jeltsch , J. M. , Krozowski , Z. , Qurin‐Stricker , C. , Gronemeyer , H. , Simpson , R. J. , Garnier , J. M. , Krust , A. , Jacob , F. and Chambon , P.Cloning of the chicken progesterone receptor . Proc. Natl. Acad. Sci. USA , 83 , 5424 – 5428 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 14. ) Savouret , J. F. , Misrahi , M. and Milgrom , E.Molecular action of progesterone . Int. J. Biochem. , 22 , 579 – 594 ( 1990. ). [DOI] [PubMed] [Google Scholar]
- 15. ) Kastner , P. , Krust , A. , Turcotte , B. , Stropp , U. , Tora , L. , Gronemeyer , H. and Chambon , P.Two distinct estrogen‐regulated promoters generate transcripts encoding the two functionally different human progesterone receptor forms A and B . EMBOJ. , 9 , 1603 – 1614 ( 1990. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 16. ) Wen , D. X. , Xu , Y. F. , Mais , D. E. , Godman , M. E. and McDonnell , D. P.The A and B isoforms of the human progesterone receptor operate through distinct signaling pathways within target cells . Mol. Cell. Biol. , 14 , 8356 – 8364 ( 1994. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 17. ) Giangrande , P. H. , Pollio , G. and McDonnell , D. P.Mapping and characterization of the functional domains responsible for the differential activity of the A and B isoforms of the human progesterone receptor . J. Biol. Chem. , 272 , 32889 – 32900 ( 1997. ). [DOI] [PubMed] [Google Scholar]
- 18. ) Vegeto , E. , Shahbaz , M. M. , Wen , D. X. , Goldman , M. E. , O'Malley , B. W. and McDonell , D. P.Human progesterone receptor A form is a cell‐and promoter‐specific repressor of human progesterone receptor B function . Mol. Endocrinol. , 7 , 1244 – 1255 ( 1993. ). [DOI] [PubMed] [Google Scholar]
- 19. ) Kraus , W. L. , Weis , K. E. and Katzenellenbogen , B. S.Inhibitory cross‐talk between steroid hormone receptors: differential targeting of estrogen receptor in the repression of its transcriptional activity by agonist‐and antagonist‐occupied progestin receptors . Mol. Cell. Biol. , 15 , 1847 – 1857 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 20. ) Clinton , G. M. and Hua , W.Estrogen action in human ovarian cancer . Crit. Rev. Oncol. Hematol. , 25 , 1 – 9 ( 1997. ). [DOI] [PubMed] [Google Scholar]
- 21. ) Kommoss , F. In “ Hormone‐Dependent Cancer ,” ed. Pasqualini J. R. and Katzenellenbogen, B. S. pp. 541 – 572 ( 1995. ). Dekker; , New York . [Google Scholar]
- 22. ) Dietel , M. and Hauptmann , S.Serous tumors of low malignant potential of the ovary: 1. Diagnostic pathology . Virchows Arch. , 436 , 403 – 412 ( 2000. ). [DOI] [PubMed] [Google Scholar]
- 23. ) Graham , J. D. and Clarke , C. L.Physiological action of progesterone in target tissues . Endocr. Rev. , 18 , 502 – 519 ( 1997. ). [DOI] [PubMed] [Google Scholar]
- 24. ) Hamilton , T. C. , Young , R. C. , McKoy , W. M. , Grotzinger , K. R. , Green , J. A. , Chu , E. W. , Whang‐Peng , J. , Rogan , A. M. , Green , W. R. and Ozols , R. F.Characterization of a human ovarian carcinoma cell line (NTH: OVCAR–3) with androgen and estrogen receptors . Cancer Res. , 43 , 5379 – 5389 ( 1983. ). [PubMed] [Google Scholar]
- 25. ) Lounis , H. , Provencher , D. , Godbout , C. , Fink , D. , Milot , M. J. and Mes‐Masson , A. M.Primary cultures of normal and tumoral human ovarian epithelium: a powerful tool for basic molecular studies . Exp. Cell Res. , 215 , 303 – 309 ( 1994. ). [DOI] [PubMed] [Google Scholar]
- 26. ) Clarke , C. L. , Zaino , P. D. , Feil , P. D. , Miller , J. V. , Steck , M. E. , Ohlsson‐Wilhelm , B. M. and Satyaswaroop , P. G.Monoclonal antibodies to human progesterone receptor: characterization by biochemical and immunohistochemical techniques . Endocrinology , 121 , 1123 – 1132 ( 1987. ). [DOI] [PubMed] [Google Scholar]
- 27. ) Mote , P. A. , Balleine , R. L. , McGowan , E. M. and Clarke , C. L.Colocalization of progesterone receptors A and B by dual immunofluorescent histochemistry in human endometrium during the menstrual cycle . J. Clin. Endocrinol. Metabol. , 84 , 2963 – 2971 ( 1999. ). [DOI] [PubMed] [Google Scholar]
- 28. ) Akahira , J. , Suzuki , T. , Ito , K. , Darnel , A. D. , Moriya , T. , Sato , S. , Yaegashi , N. , Okamura , K. and Sasano , H.Expression of 5ã‐reductases in human epithelial ovarian cancer: its correlation with androgen receptor status . Jpn. J. Cancer Res. , 92 , 926 – 932 ( 2001. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 29. ) McCarty , K. S. , Jr. , Miller , L. S. , Cox , E. B. , Konrath , J. and McCarty , K. S. , Sr.Estrogen receptor analysis . Arch. Pathol. Lab. Med. , 109 , 716 – 721 ( 1985. ). [PubMed] [Google Scholar]
- 30. ) Zeimet , A. G. , Muller‐Holzner , E. , Marth , C. and Daxenbichler , G.Immunocytochemical versus biochemical receptor determination in normal and tumorous tissues of the female reproductive tract and the breast . J. Steroid Biochem. Mol. Biol. , 49 , 365 – 372 ( 1994. ). [DOI] [PubMed] [Google Scholar]
- 31. ) Noguchi , T. and Kitawaki , J.Relationship between aromatase activity and steroid receptor levels in ovarian tumors from postmenopausal women . J. Steroid Biochem. Mol. Biol. , 44 , 657 – 660 ( 1993. ). [DOI] [PubMed] [Google Scholar]
- 32. ) Akahira , J. , Inoue , T. , Suzuki , T. , Ito , K. , Konno , R. , Sato , S. , Moriya , T. , Okamura , K. , Yajima , A. and Sasano , H.Progesterone receptor isoforms A and B in human epithelial ovarian carcinoma: immunohistochemical and RT‐PCR studies . Br. J. Cancer , 83 , 1488 – 1494 ( 2000. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 33. ) Langdon , S. P. , Gabra , H. , Bartlett , J. M. S. , Rabiaz , G. J. , Hawkins , R. A. , Tesdale , A. L. , Ritchie , A. A. , Miller , W. R. and Smyth , J. F.Functionality of the progesterone receptor in ovarian cancer and its regulation by estrogen . Clin. Cancer Res. , 4 , 2245 – 2251 ( 1998. ). [PubMed] [Google Scholar]
- 34. ) Hempling , R. E. , Piver , M. S. , Eltabbakh , G. H. and Recio , F. O.Progesterone receptor status is a significant prognostic variable of progression‐free survival in advanced epithelial ovarian cancer . Am. J. Clin. Oncol. , 21 , 447 – 451 ( 1998. ). [DOI] [PubMed] [Google Scholar]
- 35. ) Lau , K. M. , Mok , S. C. and Ho , S. M.Expression of human estrogen receptor‐α and β, progesterone receptor, and androgen receptor mRNA in normal and malignant ovarian epithelial cells . Proc. Natl. Acad. Sci. USA , 96 , 5722 – 5727 ( 1999. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 36. ) Syvala , H. , Vienonen , A. , Ylikomi , T. , Blauer , M. , Zhuang , Y. H. and Tuohimaa , P.Expression of the chicken progesterone receptor forms A and B is differentially regulated by estrogen in vivo . Biochem. Biophys. Res. Commun. , 231 , 573 – 576 ( 1997. ). [DOI] [PubMed] [Google Scholar]
- 37. ) Graham , J. D. , Roman , S. D. , McGowan , E. , Sutherland , R. L. and Clarke , C. L.Preferential stimulation of human progesterone receptor B expression by estrogen in T–47D human breast cancer cells . J. Biol. Chem. , 270 , 30693 – 30700 ( 1995. ). [DOI] [PubMed] [Google Scholar]
- 38. ) Mangal , R. K. , Wiehle , R. D. , Poindexter , A. N. and Weigel , N. L.Differential expression of uterine progesterone receptor forms A and B during the menstrual cycle . J. Steroid Biochem. Mol. Biol. , 63 , 195 – 202 ( 1997. ). [DOI] [PubMed] [Google Scholar]
- 39. ) Kauppila , A.Progestin therapy of endometrial, breast and ovarian carcinoma: a review of clinical observations . Acta Obstet. Gynecol. Scand. , 63 , 441 – 450 ( 1984. ). [DOI] [PubMed] [Google Scholar]
- 40. ) Ahlgren , J. D. , Ellison , N. M. , Gottlieb , R. J. , Laluna , F. , Lokich , J. J. , Sinclair , P. R. , Ueno , W. , Wampler , G. L. , Yeung , K. Y. , Alt , D. and Fryer , J. G.Hormonal palliation of chemoresistant ovarian cancer: three consecutive Phase II trials of the Mid‐Atlantic Oncology Program . J. Clin. Oncol. , 11 , 1957 – 1968 ( 1993. ). [DOI] [PubMed] [Google Scholar]
- 41. ) Graham , J. D. , Yeates , C. , Balleine , R. L. , Harvey , S. S. , Milliken , J. S. , Bilous , A. M. and Clarke , C. L.Characterization of progesterone receptor A and B proteins in human breast cancer . Cancer Res. , 55 , 5063 – 5068 ( 1995. ). [PubMed] [Google Scholar]
- 42. ) Graham , J. D. , Yeates , C. , Balleine , R. L. , Harvey , S. S. , Milliken , J. S. , Bilous , A. M. and Clarke , C. L.Progesterone receptor A and B protein expression in human breast cancer . J. Steroid Biochem. Mol. Biol , 56 , 93 – 98 ( 1996. ). [DOI] [PubMed] [Google Scholar]
- 43. ) Ariga , N. , Suzuki , T. , Moriya , T. , Kimura , M. , Inoue , T. , Ohuchi , N. and Sasano , H.Progesterone receptor A and B isoforms in the human breast and its disorders . Jpn. J. Cancer Res. , 92 , 302 – 308 ( 2001. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 44. ) Kumar , N. S. , Richer , J. , Owen , G. , Litman , E. , Horwitz , K. B. and Leslie , K. K.Selective down‐regulation of progesterone receptor isoforms B in poorly differentiated human endometrial cancer cells: implications for unopposed estrogen action . Cancer Res. , 58 , 1860 – 1865 ( 1998. ). [PubMed] [Google Scholar]
- 45. ) Sasaki , M. , Dharia , A. , Oh , B. R. , Tanaka , Y. , Fujimoto , S. and Dahiya , D.Progesterone receptor B gene inactivation and CpG hypermethylation in human uterine endometrial cancer . Cancer Res. , 61 , 97 – 102 ( 2001. ). [PubMed] [Google Scholar]
- 46. ) Mulac‐Jericevic , B. , Mullinax , R. A. , De Mayo , F. J. , Lydon , J. P. and Conneely , O. M.Subgroup of reproductive functions of progesterone mediated by progesterone receptor‐B isoform . Science , 289 , 1751 – 1754 ( 2000. ). [DOI] [PubMed] [Google Scholar]