Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
BMC Cancer
Published in: BMC Cancer 1/2006

Open Access 01-12-2006 | Research article

Effects of estradiol and medroxyprogesterone acetate on morphology, proliferation and apoptosis of human breast tissue in organ cultures

Authors: Natalija Eigėlienė, Pirkko Härkönen, Risto Erkkola

Published in: BMC Cancer | Issue 1/2006

Login to get access

Abstract

Background

Human breast tissue undergoes phases of proliferation, differentiation and regression regulated by changes of the levels of circulating sex hormones during the menstrual cycle or aging. Ovarian hormones also likely play a key role in the etiology and biology of breast cancer. Reports concerning the proliferative effects of steroid hormones on the normal epithelium of human breast have been conflicting. Some studies have shown that steroid hormones may predispose breast epithelial cells to malignant changes by stimulating their proliferation, which is known to be regulated tightly by stromal cells.
The aim of this study was to investigate the effects of 17β-estradiol and medroxyprogesterone acetate on proliferation, apoptosis, expression of differentiation markers and steroid hormone receptors in breast epithelium using an in vitro model of freshly isolated human breast tissue, in which a proper interaction of breast epithelium and stroma has been maintained.

Methods

Human breast tissues were obtained from women undergoing surgery for breast tumours. Peritumoral tissues were excised and explants were cultured for 3 weeks in medium supplemented with E2 or MPA or with E2+MPA. Endpoints included histopathological, histomorphometric and immunohistochemical assessment of the breast explants.

Results

Culture of breast explants for 14 or 21 days with steroid hormones increased proliferative activity and the thickness of acinar and ductal epithelium. E2-treatment led to hyperplastic epithelial morphology, MPA to hypersecretory single-layered epithelium and E2+MPA to multilayered but organised epithelium.
The proliferative response to E2 in comparison to control (p < 0.001) was more pronounced than to MPA (p < 0.05) or E2+MPA (p < 0.05) at 7 and 14 days for Ki-67 and PCNA. E2 treatment also decreased the proportion of apoptotic cells after 7 (p < 0.01) and 14 (p < 0.01) days. In addition, the relative number of ERα, ERβ and PR positive epithelial cells was decreased by all hormonal treatments.

Conclusion

Organ culture system provides a model for studying the direct effects of steroid hormones and their analogues on postmenopausal human breast tissue.
Addition of E2 or MPA or E2+MPA to breast explants caused characteristic changes in morphology, stimulated epithelial proliferation, lowered apoptosis ratio and decreased the relative number of epithelial cells expressing ERα, ERβ and PR.
Appendix
Available only for authorised users
Literature
1.
Black RJ, Bray F, Ferlay J, Parkin DM: Cancer incidence and mortality in the European Union: cancer registry data and estimates of national incidence for 1990. Eur J Cancer. 1997, 33 (7): 1075-1107. 10.1016/S0959-8049(96)00492-3.CrossRefPubMed
2.
Grady D, Rubin SM, Petitti DB, Fox CS, Black D, Ettinger B, Ernster VL, Cummings SR: Hormone therapy to prevent disease and prolong life in postmenopausal women. Ann Intern Med. 1992, 117 (12): 1016-1037.CrossRefPubMed
3.
Henderson BE, Feigelson HS: Hormonal carcinogenesis. Carcinogenesis. 2000, 21 (3): 427-433. 10.1093/carcin/21.3.427.CrossRefPubMed
4.
Preston-Martin S, Pike MC, Ross RK, Jones PA, Henderson BE: Increased cell division as a cause of human cancer. Cancer Res. 1990, 50 (23): 7415-7421.PubMed
5.
Feigelson HS, Ross RK, Yu MC, Coetzee GA, Reichardt JK, Henderson BE: Sex steroid hormones and genetic susceptibility to breast and prostate cancer. Drug Metab Rev. 1998, 30 (2): 421-434.CrossRefPubMed
6.
Rossouw JE, Anderson GL, Prentice RL, LaCroix AZ, Kooperberg C, Stefanick ML, Jackson RD, Beresford SA, Howard BV, Johnson KC, Kotchen JM, Ockene J: Risks and benefits of estrogen plus progestin in healthy postmenopausal women: principal results From the Women's Health Initiative randomized controlled trial. Jama. 2002, 288 (3): 321-333. 10.1001/jama.288.3.321.CrossRefPubMed
7.
Breast cancer and hormone replacement therapy: collaborative reanalysis of data from 51 epidemiological studies of 52,705 women with breast cancer and 108,411 women without breast cancer. Collaborative Group on Hormonal Factors in Breast Cancer. Lancet. 1997, 350 (9084): 1047-1059. 10.1016/S0140-6736(97)08233-0.
8.
Stefanick ML, Anderson GL, Margolis KL, Hendrix SL, Rodabough RJ, Paskett ED, Lane DS, Hubbell FA, Assaf AR, Sarto GE, Schenken RS, Yasmeen S, Lessin L, Chlebowski RT: Effects of conjugated equine estrogens on breast cancer and mammography screening in postmenopausal women with hysterectomy. Jama. 2006, 295 (14): 1647-1657. 10.1001/jama.295.14.1647.CrossRefPubMed
9.
Bergkvist L, Persson I: Hormone replacement therapy and breast cancer. A review of current knowledge. Drug Saf. 1996, 15 (5): 360-370.CrossRefPubMed
10.
Soderqvist G, Isaksson E, von Schoultz B, Carlstrom K, Tani E, Skoog L: Proliferation of breast epithelial cells in healthy women during the menstrual cycle. Am J Obstet Gynecol. 1997, 176 (1 Pt 1): 123-128. 10.1016/S0002-9378(97)80024-5.CrossRefPubMed
11.
Olsson H, Jernstrom H, Alm P, Kreipe H, Ingvar C, Jonsson PE, Ryden S: Proliferation of the breast epithelium in relation to menstrual cycle phase, hormonal use, and reproductive factors. Breast Cancer Res Treat. 1996, 40 (2): 187-196. 10.1007/BF01806214.CrossRefPubMed
12.
Meyer JS: Cell proliferation in normal human breast ducts, fibroadenomas, and other ductal hyperplasias measured by nuclear labeling with tritiated thymidine. Effects of menstrual phase, age, and oral contraceptive hormones. Hum Pathol. 1977, 8 (1): 67-81.CrossRefPubMed
13.
Potten CS, Watson RJ, Williams GT, Tickle S, Roberts SA, Harris M, Howell A: The effect of age and menstrual cycle upon proliferative activity of the normal human breast. Br J Cancer. 1988, 58 (2): 163-170.CrossRefPubMedPubMedCentral
14.
Going JJ, Anderson TJ, Battersby S, MacIntyre CC: Proliferative and secretory activity in human breast during natural and artificial menstrual cycles. Am J Pathol. 1988, 130 (1): 193-204.PubMedPubMedCentral
15.
Kramer EA, Seeger H, Kramer B, Wallwiener D, Mueck AO: The effects of progesterone, medroxyprogesterone acetate, and norethisterone on growth factor- and estradiol-treated human cancerous and noncancerous breast cells. Menopause. 2005, 12 (4): 468-474. 10.1097/01.GME.0000155206.53856.41.CrossRefPubMed
16.
Zhuang YH, Saaristo R, Ylikomi T: An in vitro long-term culture model for normal human mammary gland: expression and regulation of steroid receptors. Cell Tissue Res. 2003, 311 (2): 217-226.PubMed
17.
18.
Stute P, Wood CE, Kaplan JR, Cline JM: Cyclic changes in the mammary gland of cynomolgus macaques. Fertil Steril. 2004, 82 Suppl 3: 1160-1170. 10.1016/j.fertnstert.2004.04.035.CrossRefPubMed
19.
Isaksson E, Wang H, Sahlin L, von Schoultz B, Cline JM, von Schoultz E: Effects of long-term HRT and tamoxifen on the expression of progesterone receptors A and B in breast tissue from surgically postmenopausal cynomolgus macaques. Breast Cancer Res Treat. 2003, 79 (2): 233-239. 10.1023/A:1023925906199.CrossRefPubMed
20.
Cline JM, Soderqvist G, von Schoultz E, Skoog L, von Schoultz B: Effects of conjugated estrogens, medroxyprogesterone acetate, and tamoxifen on the mammary glands of macaques. Breast Cancer Res Treat. 1998, 48 (3): 221-229. 10.1023/A:1005984932268.CrossRefPubMed
21.
Levine JF, Stockdale FE: Cell-cell interactions promote mammary epithelial cell differentiation. J Cell Biol. 1985, 100 (5): 1415-1422. 10.1083/jcb.100.5.1415.CrossRefPubMed
22.
Gache C, Berthois Y, Martin PM, Saez S: Positive regulation of normal and tumoral mammary epithelial cell proliferation by fibroblasts in coculture. In Vitro Cell Dev Biol Anim. 1998, 34 (4): 347-351.CrossRefPubMed
23.
Ronnov-Jessen L, Petersen OW, Bissell MJ: Cellular changes involved in conversion of normal to malignant breast: importance of the stromal reaction. Physiol Rev. 1996, 76 (1): 69-125.PubMed
24.
Nevalainen MT, Harkonen PL, Valve EM, Ping W, Nurmi M, Martikainen PM: Hormone regulation of human prostate in organ culture. Cancer Res. 1993, 53 (21): 5199-5207.PubMed
25.
Nevalainen MT, Valve EM, Makela SI, Blauer M, Tuohimaa PJ, Harkonen PL: Estrogen and prolactin regulation of rat dorsal and lateral prostate in organ culture. Endocrinology. 1991, 129 (2): 612-622.CrossRefPubMed
26.
Trowell OA: The culture of mature organs in a synthetic medium. Exp Cell Res. 1959, 16 (1): 118-147. 10.1016/0014-4827(59)90201-0.CrossRefPubMed
27.
Hillman EA, Vocci MJ, Combs JW, Sanefuji H, Robbins T, Janss DH, Harris CC, Trump BF: Human breast organ culture studies. Methods Cell Biol. 1980, 21B: 79-106.CrossRefPubMed
28.
Hillman EA, Valerio MG, Halter SA, Barrett-Boone LA, Trump BF: Long-term explant culture of normal mammary epithelium. Cancer Res. 1983, 43 (1): 245-257.PubMed
29.
30.
Sadi MV, Barrack ER: Determination of growth fraction in advanced prostate cancer by Ki-67 immunostaining and its relationship to the time to tumor progression after hormonal therapy. Cancer. 1991, 67 (12): 3065-3071. 10.1002/1097-0142(19910615)67:12<3065::AID-CNCR2820671222>3.0.CO;2-U.CrossRefPubMed
31.
Hofseth LJ, Raafat AM, Osuch JR, Pathak DR, Slomski CA, Haslam SZ: Hormone replacement therapy with estrogen or estrogen plus medroxyprogesterone acetate is associated with increased epithelial proliferation in the normal postmenopausal breast. J Clin Endocrinol Metab. 1999, 84 (12): 4559-4565. 10.1210/jc.84.12.4559.PubMed
32.
Thike AA, Chng MJ, Fook-Chong S, Tan PH: Immunohistochemical expression of hormone receptors in invasive breast carcinoma: correlation of results of H-score with pathological parameters. Pathology. 2001, 33 (1): 21-25. 10.1080/00313020120034858.CrossRefPubMed
33.
Mourits MJ, Ten Hoor KA, van der Zee AG, Willemse PH, de Vries EG, Hollema H: The effects of tamoxifen on proliferation and steroid receptor expression in postmenopausal endometrium. J Clin Pathol. 2002, 55 (7): 514-519.CrossRefPubMedPubMedCentral
34.
Yu CC, Filipe MI: Update on proliferation-associated antibodies applicable to formalin-fixed paraffin-embedded tissue and their clinical applications. Histochem J. 1993, 25 (12): 843-853.CrossRefPubMed
35.
Foidart JM, Colin C, Denoo X, Desreux J, Beliard A, Fournier S, de Lignieres B: Estradiol and progesterone regulate the proliferation of human breast epithelial cells. Fertil Steril. 1998, 69 (5): 963-969. 10.1016/S0015-0282(98)00042-9.CrossRefPubMed
36.
Laidlaw IJ, Clarke RB, Howell A, Owen AW, Potten CS, Anderson E: The proliferation of normal human breast tissue implanted into athymic nude mice is stimulated by estrogen but not progesterone. Endocrinology. 1995, 136 (1): 164-171. 10.1210/en.136.1.164.PubMed
37.
Clarke RB, Howell A, Anderson E: Estrogen sensitivity of normal human breast tissue in vivo and implanted into athymic nude mice: analysis of the relationship between estrogen-induced proliferation and progesterone receptor expression. Breast Cancer Res Treat. 1997, 45 (2): 121-133. 10.1023/A:1005805831460.CrossRefPubMed
38.
Kuhl H: Pharmacology of estrogens and progestogens: influence of different routes of administration. Climacteric. 2005, 8 Suppl 1: 3-63.CrossRefPubMed
39.
Russo J, Russo IH: Development of the human breast. Maturitas. 2004, 49 (1): 2-15. 10.1016/j.maturitas.2004.04.011.CrossRefPubMed
40.
Parmar H, Cunha GR: Epithelial-stromal interactions in the mouse and human mammary gland in vivo. Endocr Relat Cancer. 2004, 11 (3): 437-458. 10.1677/erc.1.00659.CrossRefPubMed
41.
Russo IH, Russo J: Role of hormones in mammary cancer initiation and progression. J Mammary Gland Biol Neoplasia. 1998, 3 (1): 49-61. 10.1023/A:1018770218022.CrossRefPubMed
42.
Berstein LM, Tsyrlina EV, Kolesnik OS, Gamajunova VB, Adlercreutz H: Catecholestrogens excretion in smoking and non-smoking postmenopausal women receiving estrogen replacement therapy. J Steroid Biochem Mol Biol. 2000, 72 (3-4): 143-147. 10.1016/S0960-0760(00)00038-8.CrossRefPubMed
43.
Seeger H, Wallwiener D, Kraemer E, Mueck AO: Comparison of possible carcinogenic estradiol metabolites: Effects on proliferation, apoptosis and metastasis of human breast cancer cells. Maturitas. 2006, 54 (1): 72-77. 10.1016/j.maturitas.2005.08.010.CrossRefPubMed
44.
Russo J, Ao X, Grill C, Russo IH: Pattern of distribution of cells positive for estrogen receptor alpha and progesterone receptor in relation to proliferating cells in the mammary gland. Breast Cancer Res Treat. 1999, 53 (3): 217-227. 10.1023/A:1006186719322.CrossRefPubMed
45.
Kuiper GG, Enmark E, Pelto-Huikko M, Nilsson S, Gustafsson JA: Cloning of a novel receptor expressed in rat prostate and ovary. Proc Natl Acad Sci U S A. 1996, 93 (12): 5925-5930. 10.1073/pnas.93.12.5925.CrossRefPubMedPubMedCentral
46.
Speirs V, Skliris GP, Burdall SE, Carder PJ: Distinct expression patterns of ER alpha and ER beta in normal human mammary gland. J Clin Pathol. 2002, 55 (5): 371-374.CrossRefPubMedPubMedCentral
47.
Skliris GP, Parkes AT, Limer JL, Burdall SE, Carder PJ, Speirs V: Evaluation of seven oestrogen receptor beta antibodies for immunohistochemistry, western blotting, and flow cytometry in human breast tissue. J Pathol. 2002, 197 (2): 155-162. 10.1002/path.1077.CrossRefPubMed
48.
Clarke RB, Howell A, Potten CS, Anderson E: Dissociation between steroid receptor expression and cell proliferation in the human breast. Cancer Res. 1997, 57 (22): 4987-4991.PubMed
49.
Anderson E, Clarke RB, Howell A: Estrogen responsiveness and control of normal human breast proliferation. J Mammary Gland Biol Neoplasia. 1998, 3 (1): 23-35. 10.1023/A:1018718117113.CrossRefPubMed
50.
Anderson E: The role of oestrogen and progesterone receptors in human mammary development and tumorigenesis. Breast Cancer Res. 2002, 4 (5): 197-201. 10.1186/bcr452.CrossRefPubMedPubMedCentral
51.
Cheng G, Li Y, Omoto Y, Wang Y, Berg T, Nord M, Vihko P, Warner M, Piao YS, Gustafsson JA: Differential regulation of estrogen receptor (ER)alpha and ERbeta in primate mammary gland. J Clin Endocrinol Metab. 2005, 90 (1): 435-444. 10.1210/jc.2004-0861.CrossRefPubMed
52.
Gustafsson JA: ERbeta scientific visions translate to clinical uses. Climacteric. 2006, 9 (3): 156-160. 10.1080/14689360600734328.CrossRefPubMed
53.
Herynk MH, Fuqua SA: Estrogen receptor mutations in human disease. Endocr Rev. 2004, 25 (6): 869-898. 10.1210/er.2003-0010.CrossRefPubMed
54.
Conneely OM, Jericevic BM, Lydon JP: Progesterone receptors in mammary gland development and tumorigenesis. J Mammary Gland Biol Neoplasia. 2003, 8 (2): 205-214. 10.1023/A:1025952924864.CrossRefPubMed
55.
Graham JD, Clarke CL: Physiological action of progesterone in target tissues. Endocr Rev. 1997, 18 (4): 502-519. 10.1210/er.18.4.502.PubMed
56.
Anderson E, Clarke RB: Steroid receptors and cell cycle in normal mammary epithelium. J Mammary Gland Biol Neoplasia. 2004, 9 (1): 3-13. 10.1023/B:JOMG.0000023584.01750.16.CrossRefPubMed
57.
Gustafsson JA, Warner M: Estrogen receptor beta in the breast: role in estrogen responsiveness and development of breast cancer. J Steroid Biochem Mol Biol. 2000, 74 (5): 245-248. 10.1016/S0960-0760(00)00130-8.CrossRefPubMed
Metadata
Title
Effects of estradiol and medroxyprogesterone acetate on morphology, proliferation and apoptosis of human breast tissue in organ cultures
Authors
Natalija Eigėlienė
Pirkko Härkönen
Risto Erkkola
Publication date
01-12-2006
Publisher
BioMed Central
Published in
BMC Cancer / Issue 1/2006
Electronic ISSN: 1471-2407
DOI
https://doi.org/10.1186/1471-2407-6-246

Other articles of this Issue 1/2006

BMC Cancer 1/2006 Go to the issue

Research article

The associated expression of Maspin and Bax proteins as a potential prognostic factor in intrahepatic cholangiocarcinoma

Research article

Leptin and leptin receptor polymorphisms are associated with increased risk and poor prognosis of breast carcinoma

Case report

Spontaneous regression of bone metastasis from renal cell carcinoma; A case report

Research article

Prognostic significance of fascin expression in advanced colorectal cancer: an immunohistochemical study of colorectal adenomas and adenocarcinomas

Research article

An association of a simultaneous nuclear and cytoplasmic localization of Fra-1 with breast malignancy

Research article

Zoledronic acid treatment impairs protein geranyl-geranylation for biological effects in prostatic cells
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits