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Steroid hormone secretion in inflammatory breast cancer cell lines

  • Juan Carlos Illera EMAIL logo , Sara Caceres , Laura Peña , Paloma J. de Andres , Beatriz Monsalve , Maria J. Illera , Wendy A. Woodward , James M. Reuben and Gema Silvan
Published/Copyright: October 23, 2015

Abstract

Inflammatory breast carcinoma (IBC) is a special type of breast cancer with a poor survival rate. Though several IBC cell lines have been established, recently a first IMC cell line was established. The aims of this study were: (1) to validate a highly sensitive, reliable, accurate and direct amplified enzyme immunoassay (EIA) to measure several cell-secreted steroid hormones: progesterone (P4), androstenedione (A4), testosterone (T), 17β-estradiol (E2) and estrone sulfate (SO4E1) in the culture medium. (2) To assess whether hormone production profile by IPC-366 cells validates the IMC model for human IBC. We validated a non-competitive amplified EIA for inflammatory breast cancer cell lines based on the results of accuracy, precision, sensitivity and parallelism. The low detection limits of the technique were: P4=13.2 pg/well, A4=2.3 pg/well, T=11.4 pg/well, E2=1.9 pg/well and SO4E1=4.5 pg/well. Intra- and inter-assay coefficient of variation percentages were <10%. The mean recovery rate of hormone added to the culture medium was >90%. In all hormones studied SUM149 have higher levels (1.4 times, but not significant) than IPC-366, and the correlation index between SUM149 and IPC-366 concentrations were >97%. We can coclude that cells of both cell lines, IPC-366 and SUM149, are capable to produce steroid hormone in culture media. The presented EIA methodology is very valuable for the detection of steroid production in culture media and could be used in hormone regulation studies and therapeutic agents in cell lines of inflammatory and non-inflammatory mammary carcinoma or other cancer cell lines in preclinical studies.


Corresponding author: Juan Carlos Illera, DVM, PhD, ECAR diplomat, MLAS, Dpto. Fisiologia Animal, Facultad de Veterinaria, Universidad Complutense de Madrid, Avda. Puerta de Hierro s/n, 28040 Madrid, Spain, Phone: +913943867, E-mail: ; and Department of Animal Physiology, Surgery and Pathology, School of Veterinary Medicine, Complutense University of Madrid (UCM), Spain

Acknowledgments

This research was supported by the Spanish Ministry of Science and Education (research project No SAF 2009-10572).

Conflict of interest statement: The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.

References

1. Levine PH, Steinhorn SC, Ries LG, Aron JL. Inflammatory breast cancer. The experience of the surveillance, epidemiology, and end results (SEER) program. J Nat Cancer Inst 1985;74:291–7.Search in Google Scholar

2. Somlo G, Doroshow JH, Forman SJ, Odom-Mayon T, Lee J, Chow W, Hamasaki V, Leong L, Morgan Jr R, Margolin K, Raschko J, Shibata S, Telef M, Yen Y, Simpson J, Molina A. High-dose chemotherapy and stem-cell rescue in the treatment of high-risk breast cancer: prognostic indicators of progression-free and overall survival. J Clin Oncol 1997;15:2882–93.10.1200/JCO.1997.15.8.2882Search in Google Scholar

3. Victor H, Horwitz EM, Kini VR, Martinez AA, Pettinga JE, Dmuchowski CF, Decker DA, Wilner FM, Vicini FA. Impact of clinical, pathologic, and treatment-related factors of patients with local advanced breast cancer treated with multimodality therapy. Am J Clin Oncol 1999;22:119–25.10.1097/00000421-199904000-00003Search in Google Scholar

4. Peña L, Silván G, Pérez-Alenza MD, Nieto A, Illera JC. Steroid hormone profile of canine inflammatory mammary carcinoma: a preliminary study. J Steroid Biochem Mol Biol 2003;84:211–6.10.1016/S0960-0760(03)00030-XSearch in Google Scholar

5. Van Golen KJ, Davies S, Wu ZF, Wang Y, Bucana CD, Root H, Chandrasekharappa S, Strawderman M, Ethier SP, Merajver SD. A novel putative low-affinity insulin-like growth factor-binding protein, LIBC (lost in inflammatory breast cancer), and RhoC GTPasa correlate with the inflammatory breast cancer phenotype. Clin Cancer Res 1999;5:5832–8.Search in Google Scholar

6. Caceres S, Peña L, de Andres PJ, Illera MJ, Lopez MS, Woodward WA, Reuben JM, Illera JC. Establishment and characterization of a new cell line of canine inflammatory mammary cancer: IPC-366. Plos One 2015;10:e0122277.10.1371/journal.pone.0122277Search in Google Scholar

7. Tavassoli FA. Inflammatory carcinoma. Infiltrating carcinoma: special types. Pathology of the Breast, 2nd ed. New York: McGraw-Hill, 1999:519–41.Search in Google Scholar

8. Giordano SH, Hortobagyi GN. Inflammatory breast cancer: clinical progress and the main problems that must be addressed. Breast Cancer Res 2003;5:284–8.10.1186/bcr608Search in Google Scholar

9. Blankenstein MA, van de Ven J, Maitimu-Smeele I, Donker GH, de Jong PC, Daroszewski J, Szymczak J, Milewicz A, Thijseen JH. Intratumoral levels of estrogens in breast cancer. J Steroid Biochem Mol Biol 1999;69:293–7.10.1016/S0960-0760(99)00048-5Search in Google Scholar

10. Abul-Hajj YJ, Iverson R, Kiang DT. Aromatization of androgens by human breast cancer. Steroids 1979;33:205–22.10.1016/0039-128X(79)90027-8Search in Google Scholar

11. Lipton A, Santner SJ, Santen RJ, Harvey HA, Feil PD, White-Hershey D, Bartholomew MJ, Antle CE. Aromatase activity in primary and metastatic human breast cancer. Cancer 1987;59:779–82.10.1002/1097-0142(19870215)59:4<779::AID-CNCR2820590419>3.0.CO;2-USearch in Google Scholar

12. Dao TL, Hayes C, Libby CPR. Steroid sulfatase activities in human breast tumors. Proc Soc Experim Biol Med 1984;146:381–4.10.3181/00379727-146-38109Search in Google Scholar

13. Pasqualini JR, Gelly C, Nguyen BL, Vella C. Importance of estrogen sulfates in breast cancer. J Steroid Biochem 1989;34:155–63.10.1016/0022-4731(89)90077-0Search in Google Scholar

14. Secky L, Svoboda M, Klameth L, Bajna E, Hamilton G, Zeillinger R, Jäger W, Thalhammer T. The sulfatase pathway for estrogen formation: targets for the treatment and diagnosis of hormone-associated tumors. J Drug Delivery 2013. DOI: 10.1155/2013/957605.10.1155/2013/957605Search in Google Scholar

15. Edery M, Goussard J, Dehennin L, Scholler R, Reiffsteck J, Drosdowsky MA. Endogenous estradiol-17β concentration in breast tumours determined by mass fragmentography and by radioimmunoassay: relationship to receptor content. Eur J Cancer 1981;17:115–20.10.1016/0014-2964(81)90220-6Search in Google Scholar

16. Pasqualini JR, Chetrite G, Blacker C, Feinstein MC, Delalonde L, Talbi M, Maloche C. Concentrations of estrone, estradiol, and estrone sulfate and evaluation of sulfatase and aromatase activities in pre- and postmenopausal breast cancer patients. J Clin Endocrinol Metab 1996;81:1460–4.10.1210/jcem.81.4.8636351Search in Google Scholar

17. Illera JC, Lorenzo PL, Silván G, Munro CJ, Illera MJ, Illera M. Enzyme immunoassay for testosterone and androstenedione in culture maturation medium from rabbit oocytes matured in vitro. Theriogenol 1997;47:1375–88.10.1016/S0093-691X(97)00129-5Search in Google Scholar

18. Munro C, Stabenfeldt G. Development of a microtitre plate enzyme immunoassay for the determination of progesterone. J Endocrinol 1984;101:41–9.10.1677/joe.0.1010041Search in Google Scholar

19. Jones I, Madej A. A sensitive microtitre plate enzyme immunoassay of oestradiol-17β in the cow and mare. J Immunoassay 1988;9:349–65.10.1080/01971528808053221Search in Google Scholar

20. Rodbard D. Statistical quality control and routine data processing for radioimmunoassays and immunoradiometric assays. Clin Chem 1974;20:1255–70.10.1093/clinchem/20.10.1255Search in Google Scholar

21. Abraham GE. Radioimmunoassay of steroids in biological fluids. J Steroid Biochem 1975;6:261–70.10.1016/0022-4731(75)90141-7Search in Google Scholar

22. SAS/STAT Guide for Personal Computers, SAS Institute Inc., Cary, NC, 1987.Search in Google Scholar

23. Munro CJ, Lasley BL. Non-radiometric methods for immunoassay of steroid hormones. In: Albertson BD, Haseltine FP, editors. Non-radiometric assays: technology and application in polypeptide and steroid hormone detection. New York: Alan R. Liss Inc., 1988:289–329.Search in Google Scholar

24. Silvan G, Illera JC, Illera M. Determination of follicular fluid estradiol levels by enzyme-linked immunosorbent assay. Steroids 1993;58:324–9.10.1016/0039-128X(93)90092-2Search in Google Scholar

25. Carneiro GF, Liu IKM, Illera JC, Munro CJ. Enzyme immunoassay for the measurement of estrone sulphate in cryptorchidsm, stallions, and donkeys, AEEP Proceedings 1998;44:3–4.Search in Google Scholar

26. Liao DJ, Dickson RB. Roles of androgens in the development, growth, and carcinogenesis of the mammary gland. J Steroid Biochem Mol Biol 2002;80;175–89.10.1016/S0960-0760(01)00185-6Search in Google Scholar

27. Queiroga FL, Pérez-Alenza MD, Silvan G, Peña L, Lopes C, Illera JC. Role of steroid hormones and prolactin in canine mammary cancer. J Steroid Biochem Mol Biol 2005;94;181–7.10.1016/j.jsbmb.2004.12.014Search in Google Scholar PubMed

28. Illera JC, Pérez-Alenza MD, Nieto A, Jiménez MA, Silvan G, Dunner S, Peña L. Steroids and receptors in canine mammary cancer. Steroids 2006;71:541–8.10.1016/j.steroids.2005.11.007Search in Google Scholar

29. Sánchez-Archidona AR, Jiménez MA, Pérez-Alenza D, Silván G, Illera JC, Peña L, Dunner S. Steroid pathway and oestrone sulphate production in canine inflammatory mammary carcinoma. J Steroid Biochem Mol Biol 2007;104:93–9.10.1016/j.jsbmb.2007.03.014Search in Google Scholar

30. Queiroga FL, Pérez-Alenza MD, Silvan G, Peña L, Lopes CS, Illera JC. Crosstalk between GH/IGF-I axis and steroid hormones (progesterone, 17beta-estradiol) in canine mammary tumours. J Steroid Biochem Mol Biol 2008;110:76–82.10.1016/j.jsbmb.2008.02.005Search in Google Scholar

31. Queiroga FL, Pérez-Alenza D, Silvan G, Peña L, Illera JC. Positive correlation of steroid hormones and EGF in canine mammary cancer. J Steroid Biochem Mol Biol 2009;115:9–13.10.1016/j.jsbmb.2009.01.018Search in Google Scholar

32. Wiebe JP. Progesterone metabolites in breat cancer. Endocr Related Cancer 2006;13:717–38.10.1677/erc.1.01010Search in Google Scholar

33. Key TJ, Appleby PN, Reeves GK. Hormones breast cancer collaborative group. Body mass index, serum sex hormones, and breast cancer risk in postmenopausal women. J Nat Cancer Inst 2003;95:1218–26.10.1093/jnci/djg022Search in Google Scholar

34. Grace PB, Taylor JL, Low YL, Luben RN, Mulligan AA, Botting NP, Dowsett M, Welch AA, Khaw KT, Wareham NJ, Day NE, Bingham SA. Phytoestrogen concentrations in serum and spot urine as biomarkers for dietary phytoestrogen intake and their relation to breast cancer risk in European prospective investigation of cancer and nutrition-norfolk. Cancer Epidemiol Biomarkers Prev 2004;13:698–708.10.1158/1055-9965.698.13.5Search in Google Scholar

35. Sturgeon SR, Potischman N, Malone KE, Dorgan JF, Daling J, Schairer C, Brinton LA. Serum levels of sex hormones and breast cancer risk in premenopausal women: a case-control study (USA). Cancer Cause Control 2004:15;45–53.10.1023/B:CACO.0000016574.79728.11Search in Google Scholar

36. Mady EA, Ramadan EE, Ossman AA. Sex steroid hormones in serum and tissue of benign and malignant breast tumors patients. Dis Markers 200;16:151–7.10.1155/2000/305940Search in Google Scholar

37. Asseryanis E, Ruecklinger E, Hellan M, Kubista E, Singer CF. Breast cancer size in postmenopausal women is correlated with body mass index and androgen serum levels. Gynecol Endocrinol 2014;18:29–36.10.1080/09513590310001651759Search in Google Scholar

38. Billich A, Nussbaumer P, Lehr P. Stimulation of MCF-7 breast cancer cell proliferation by estrone sulphate and dehydroepiandrosterone sulphate: inhibition by novel non-steroid sulfatase inhibitors. J Steroid Biochem Mol Biol 2000;73:225–35.10.1016/S0960-0760(00)00077-7Search in Google Scholar

39. Camacho L, Peña L, González Gil A, Cáceres S, Díez L, Illera JC. Establishment and characterization of a canine xenograft model of inflammatory mammary carcinoma. Res Vet Sci 2013;95:1068–75.10.1016/j.rvsc.2013.07.017Search in Google Scholar

40. Shymala G, Chou YC, Louie SG, Guzman RC, Smith GH, Nandi S. Cellular expression of estrogen and progesterone receptors in mammary glands: regulation by hormones, development and aging. J Steroid Biochem Mol Biol 2002;80:137–48.10.1016/S0960-0760(01)00182-0Search in Google Scholar

41. Bradlow Hl, Sepkovic DW. Steroids as procarcinogenic agents. Ann New York Acad Sci 2004;1028:216–32.10.1196/annals.1322.027Search in Google Scholar

42. Suzuki T, Darnel AD, Akahira JI, Ariga N, Ogawa S, Kaneko C, Takeyama J, Moriya T, Sasano H. 5α-reductases in human breast carcinoma: posible modulator of in situ androgen actions. J Clin Endocrin Metab 2001;86:289–96.10.1210/jcem.86.5.7444Search in Google Scholar

43. Santen RJ, Manni H, Harvey H, Redmond C. Endocrine treatment of breast cancer in women. Endocr Rev 1990;11:221–65.10.1210/edrv-11-2-221Search in Google Scholar

44. Mol JA, van Garderen E, Rutterman GR, Rijnberk A. New sights in the molecular mechanism of progestin-induced proliferation of mammary epithelium: induction of the local biosynthesis of growth hormone (GH) in the mammary gland of dogs, cats and humans. J Steroid Biochem Mol Biol 1996;57:67–71.10.1016/0960-0760(95)00251-0Search in Google Scholar

45. Wiebe JP, Souter L, Zhang G. Dutasteride affects progesterone metabolizing enzyme activity/expression in human breast cancer cell lines resulting in suppression of cell proliferation and detachment. J Steroid Biochem Mol Biol 2006;93:201–8.10.1016/j.jsbmb.2004.12.003Search in Google Scholar

46. Poels LG, Jap PH, Ramaekers FF, Scheres JM, Thomas CM, Vooijs CM, Croes HJ, Mungyer G. Characterization of a hormone-producing ovarian carcinoma cell line. Gynecol Oncol 1989;32:203–14.10.1016/S0090-8258(89)80034-4Search in Google Scholar

Received: 2015-6-10
Accepted: 2015-9-4
Published Online: 2015-10-23
Published in Print: 2015-12-1

©2015 by De Gruyter

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