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Am J Pathol. Jun 1996; 148(6): 1905–1914.
PMCID: PMC1861635

Immunohistochemical analysis of retinoic acid receptor-alpha in human breast tumors: retinoic acid receptor-alpha expression correlates with proliferative activity.


Retinoids are known to prevent mammary carcinogenesis in rodents and inhibit the growth of human breast cancer cells in vitro. Previously we demonstrated that retinoid inhibition of proliferation of human breast cancer cell lines is largely mediated by retinoic acid receptor (RAR)-alpha. In this study we describe for the first time the histological distribution of RAR-alpha in 33 breast lesion specimens as determined by immunostaining with RAR-alpha antibody. Nuclear staining was observed in tumor tissue and normal portions of the breast samples. Connective tissue exhibited relative uniform staining, whereas a wide range of RAR-alpha expression was found in the epithelial tumor cells. RAR-alpha protein was expressed at significantly higher levels in tumors with greater proliferative activity as determined by immunostaining with Ki-67 antibody. This suggests that RAR-alpha expression may be altered with tumor progression. Although a positive correlation between RAR-alpha mRNA levels and estrogen receptor status of breast tumors has previously been documented, we did not find such a relationship at the protein level. As RAR-alpha plays a major role in retinoid-mediated growth inhibition of human breast cancer cell in vitro, our findings suggest that patients with highly proliferating tumors could be responsive to retinoid independently of their responsiveness to (anti)-estrogens.

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  • De Luca LM. Retinoids and their receptors in differentiation, embryogenesis, and neoplasia. FASEB J. 1991 Nov;5(14):2924–2933. [PubMed]
  • Lotan R. Effects of vitamin A and its analogs (retinoids) on normal and neoplastic cells. Biochim Biophys Acta. 1980 Mar 12;605(1):33–91. [PubMed]
  • Amos B, Lotan R. Retinoid-sensitive cells and cell lines. Methods Enzymol. 1990;190:217–225. [PubMed]
  • Castaigne S, Chomienne C, Daniel MT, Ballerini P, Berger R, Fenaux P, Degos L. All-trans retinoic acid as a differentiation therapy for acute promyelocytic leukemia. I. Clinical results. Blood. 1990 Nov 1;76(9):1704–1709. [PubMed]
  • Hong WK, Endicott J, Itri LM, Doos W, Batsakis JG, Bell R, Fofonoff S, Byers R, Atkinson EN, Vaughan C, et al. 13-cis-retinoic acid in the treatment of oral leukoplakia. N Engl J Med. 1986 Dec 11;315(24):1501–1505. [PubMed]
  • Hong WK, Lippman SM, Itri LM, Karp DD, Lee JS, Byers RM, Schantz SP, Kramer AM, Lotan R, Peters LJ, et al. Prevention of second primary tumors with isotretinoin in squamous-cell carcinoma of the head and neck. N Engl J Med. 1990 Sep 20;323(12):795–801. [PubMed]
  • Lippman SM, Kavanagh JJ, Paredes-Espinoza M, Delgadillo-Madrueño F, Paredes-Casillas P, Hong WK, Holdener E, Krakoff IH. 13-cis-retinoic acid plus interferon alpha-2a: highly active systemic therapy for squamous cell carcinoma of the cervix. J Natl Cancer Inst. 1992 Feb 19;84(4):241–245. [PubMed]
  • Lippman SM, Parkinson DR, Itri LM, Weber RS, Schantz SP, Ota DM, Schusterman MA, Krakoff IH, Gutterman JU, Hong WK. 13-cis-retinoic acid and interferon alpha-2a: effective combination therapy for advanced squamous cell carcinoma of the skin. J Natl Cancer Inst. 1992 Feb 19;84(4):235–241. [PubMed]
  • Petkovich M, Brand NJ, Krust A, Chambon P. A human retinoic acid receptor which belongs to the family of nuclear receptors. Nature. 1987 Dec 3;330(6147):444–450. [PubMed]
  • Mangelsdorf DJ, Ong ES, Dyck JA, Evans RM. Nuclear receptor that identifies a novel retinoic acid response pathway. Nature. 1990 May 17;345(6272):224–229. [PubMed]
  • Giguere V, Ong ES, Segui P, Evans RM. Identification of a receptor for the morphogen retinoic acid. Nature. 1987 Dec 17;330(6149):624–629. [PubMed]
  • de Thé H, Marchio A, Tiollais P, Dejean A. A novel steroid thyroid hormone receptor-related gene inappropriately expressed in human hepatocellular carcinoma. Nature. 1987 Dec 17;330(6149):667–670. [PubMed]
  • Brand N, Petkovich M, Krust A, Chambon P, de Thé H, Marchio A, Tiollais P, Dejean A. Identification of a second human retinoic acid receptor. Nature. 1988 Apr 28;332(6167):850–853. [PubMed]
  • Krust A, Kastner P, Petkovich M, Zelent A, Chambon P. A third human retinoic acid receptor, hRAR-gamma. Proc Natl Acad Sci U S A. 1989 Jul;86(14):5310–5314. [PMC free article] [PubMed]
  • Hamada K, Gleason SL, Levi BZ, Hirschfeld S, Appella E, Ozato K. H-2RIIBP, a member of the nuclear hormone receptor superfamily that binds to both the regulatory element of major histocompatibility class I genes and the estrogen response element. Proc Natl Acad Sci U S A. 1989 Nov;86(21):8289–8293. [PMC free article] [PubMed]
  • Mangelsdorf DJ, Borgmeyer U, Heyman RA, Zhou JY, Ong ES, Oro AE, Kakizuka A, Evans RM. Characterization of three RXR genes that mediate the action of 9-cis retinoic acid. Genes Dev. 1992 Mar;6(3):329–344. [PubMed]
  • Kastner P, Krust A, Mendelsohn C, Garnier JM, Zelent A, Leroy P, Staub A, Chambon P. Murine isoforms of retinoic acid receptor gamma with specific patterns of expression. Proc Natl Acad Sci U S A. 1990 Apr;87(7):2700–2704. [PMC free article] [PubMed]
  • Leroy P, Krust A, Zelent A, Mendelsohn C, Garnier JM, Kastner P, Dierich A, Chambon P. Multiple isoforms of the mouse retinoic acid receptor alpha are generated by alternative splicing and differential induction by retinoic acid. EMBO J. 1991 Jan;10(1):59–69. [PMC free article] [PubMed]
  • Zelent A, Mendelsohn C, Kastner P, Krust A, Garnier JM, Ruffenach F, Leroy P, Chambon P. Differentially expressed isoforms of the mouse retinoic acid receptor beta generated by usage of two promoters and alternative splicing. EMBO J. 1991 Jan;10(1):71–81. [PMC free article] [PubMed]
  • Gudas LJ. Retinoids, retinoid-responsive genes, cell differentiation, and cancer. Cell Growth Differ. 1992 Sep;3(9):655–662. [PubMed]
  • Leid M, Kastner P, Chambon P. Multiplicity generates diversity in the retinoic acid signalling pathways. Trends Biochem Sci. 1992 Oct;17(10):427–433. [PubMed]
  • Heyman RA, Mangelsdorf DJ, Dyck JA, Stein RB, Eichele G, Evans RM, Thaller C. 9-cis retinoic acid is a high affinity ligand for the retinoid X receptor. Cell. 1992 Jan 24;68(2):397–406. [PubMed]
  • Levin AA, Sturzenbecker LJ, Kazmer S, Bosakowski T, Huselton C, Allenby G, Speck J, Kratzeisen C, Rosenberger M, Lovey A, et al. 9-cis retinoic acid stereoisomer binds and activates the nuclear receptor RXR alpha. Nature. 1992 Jan 23;355(6358):359–361. [PubMed]
  • Ratko TA, Detrisac CJ, Dinger NM, Thomas CF, Kelloff GJ, Moon RC. Chemopreventive efficacy of combined retinoid and tamoxifen treatment following surgical excision of a primary mammary cancer in female rats. Cancer Res. 1989 Aug 15;49(16):4472–4476. [PubMed]
  • Moon RC, Mehta RG. Chemoprevention of mammary cancer by retinoids. Basic Life Sci. 1990;52:213–224. [PubMed]
  • Anzano MA, Byers SW, Smith JM, Peer CW, Mullen LT, Brown CC, Roberts AB, Sporn MB. Prevention of breast cancer in the rat with 9-cis-retinoic acid as a single agent and in combination with tamoxifen. Cancer Res. 1994 Sep 1;54(17):4614–4617. [PubMed]
  • Welsch CW. Host factors affecting the growth of carcinogen-induced rat mammary carcinomas: a review and tribute to Charles Brenton Huggins. Cancer Res. 1985 Aug;45(8):3415–3443. [PubMed]
  • Boccardo F, Canobbio L, Resasco M, Decensi AU, Pastorino G, Brema F. Phase II study of tamoxifen and high-dose retinyl acetate in patients with advanced breast cancer. J Cancer Res Clin Oncol. 1990;116(5):503–506. [PubMed]
  • Cobleigh MA, Dowlatshahi K, Deutsch TA, Mehta RG, Moon RC, Minn F, Benson AB, 3rd, Rademaker AW, Ashenhurst JB, Wade JL, 3rd, et al. Phase I/II trial of tamoxifen with or without fenretinide, an analog of vitamin A, in women with metastatic breast cancer. J Clin Oncol. 1993 Mar;11(3):474–477. [PubMed]
  • Costa A, Sacchini V, Decensi A. Retinoids and tamoxifen in breast cancer chemoprevention. Int J Clin Lab Res. 1993;23(2):53–55. [PubMed]
  • Marth C, Mayer I, Daxenbichler G. Effect of retinoic acid and 4-hydroxytamoxifen on human breast cancer cell lines. Biochem Pharmacol. 1984 Jul 15;33(14):2217–2221. [PubMed]
  • Fontana JA, Miranda D, Mezu AB. Retinoic acid inhibition of human breast carcinoma proliferation is accompanied by inhibition of the synthesis of a Mr 39,000 protein. Cancer Res. 1990 Apr 1;50(7):1977–1982. [PubMed]
  • van der Burg B, van der Leede BM, Kwakkenbos-Isbrücker L, Salverda S, de Laat SW, van der Saag PT. Retinoic acid resistance of estradiol-independent breast cancer cells coincides with diminished retinoic acid receptor function. Mol Cell Endocrinol. 1993 Feb;91(1-2):149–157. [PubMed]
  • Rubin M, Fenig E, Rosenauer A, Menendez-Botet C, Achkar C, Bentel JM, Yahalom J, Mendelsohn J, Miller WH., Jr 9-Cis retinoic acid inhibits growth of breast cancer cells and down-regulates estrogen receptor RNA and protein. Cancer Res. 1994 Dec 15;54(24):6549–6556. [PubMed]
  • Roman SD, Clarke CL, Hall RE, Alexander IE, Sutherland RL. Expression and regulation of retinoic acid receptors in human breast cancer cells. Cancer Res. 1992 Apr 15;52(8):2236–2242. [PubMed]
  • Roman SD, Ormandy CJ, Manning DL, Blamey RW, Nicholson RI, Sutherland RL, Clarke CL. Estradiol induction of retinoic acid receptors in human breast cancer cells. Cancer Res. 1993 Dec 15;53(24):5940–5945. [PubMed]
  • van der Leede BJ, Folkers GE, van den Brink CE, van der Saag PT, van der Burg B. Retinoic acid receptor alpha 1 isoform is induced by estradiol and confers retinoic acid sensitivity in human breast cancer cells. Mol Cell Endocrinol. 1995 Mar;109(1):77–86. [PubMed]
  • Sheikh MS, Shao ZM, Li XS, Dawson M, Jetten AM, Wu S, Conley BA, Garcia M, Rochefort H, Fontana JA. Retinoid-resistant estrogen receptor-negative human breast carcinoma cells transfected with retinoic acid receptor-alpha acquire sensitivity to growth inhibition by retinoids. J Biol Chem. 1994 Aug 26;269(34):21440–21447. [PubMed]
  • BLOOM HJ, RICHARDSON WW. Histological grading and prognosis in breast cancer; a study of 1409 cases of which 359 have been followed for 15 years. Br J Cancer. 1957 Sep;11(3):359–377. [PMC free article] [PubMed]
  • Gaub MP, Rochette-Egly C, Lutz Y, Ali S, Matthes H, Scheuer I, Chambon P. Immunodetection of multiple species of retinoic acid receptor alpha: evidence for phosphorylation. Exp Cell Res. 1992 Aug;201(2):335–346. [PubMed]
  • Charpin C, Andrac L, Vacheret H, Habib MC, Devictor B, Lavaut MN, Toga M. Multiparametric evaluation (SAMBA) of growth fraction (monoclonal Ki67) in breast carcinoma tissue sections. Cancer Res. 1988 Aug 1;48(15):4368–4374. [PubMed]
  • Pertschuk LP, Kim DS, Nayer K, Feldman JG, Eisenberg KB, Carter AC, Rong ZT, Thelmo WL, Fleisher J, Greene GL. Immunocytochemical estrogen and progestin receptor assays in breast cancer with monoclonal antibodies. Histopathologic, demographic, and biochemical correlations and relationship to endocrine response and survival. Cancer. 1990 Oct 15;66(8):1663–1670. [PubMed]
  • Reiner A, Neumeister B, Spona J, Reiner G, Schemper M, Jakesz R. Immunocytochemical localization of estrogen and progesterone receptor and prognosis in human primary breast cancer. Cancer Res. 1990 Nov 1;50(21):7057–7061. [PubMed]
  • Rudas M, Gnant MF, Mittlböck M, Neumayer R, Kummer A, Jakesz R, Reiner G, Reiner A. Thymidine labeling index and Ki-67 growth fraction in breast cancer: comparison and correlation with prognosis. Breast Cancer Res Treat. 1994;32(2):165–175. [PubMed]
  • van Gorp J, Weiping L, Jacobse K, Liu YH, Li FY, De Weger RA, Li G. Epstein-Barr virus in nasal T-cell lymphomas (polymorphic reticulosis/midline malignant reticulosis) in western China. J Pathol. 1994 Jun;173(2):81–87. [PubMed]
  • de The H, Marchio A, Tiollais P, Dejean A. Differential expression and ligand regulation of the retinoic acid receptor alpha and beta genes. EMBO J. 1989 Feb;8(2):429–433. [PMC free article] [PubMed]
  • van der Leede BM, van den Brink CE, van der Saag PT. Retinoic acid receptor and retinoid X receptor expression in retinoic acid-resistant human tumor cell lines. Mol Carcinog. 1993;8(2):112–122. [PubMed]
  • Kastner P, Perez A, Lutz Y, Rochette-Egly C, Gaub MP, Durand B, Lanotte M, Berger R, Chambon P. Structure, localization and transcriptional properties of two classes of retinoic acid receptor alpha fusion proteins in acute promyelocytic leukemia (APL): structural similarities with a new family of oncoproteins. EMBO J. 1992 Feb;11(2):629–642. [PMC free article] [PubMed]
  • Gerdes J, Lelle RJ, Pickartz H, Heidenreich W, Schwarting R, Kurtsiefer L, Stauch G, Stein H. Growth fractions in breast cancers determined in situ with monoclonal antibody Ki-67. J Clin Pathol. 1986 Sep;39(9):977–980. [PMC free article] [PubMed]
  • Potischman N, McCulloch CE, Byers T, Nemoto T, Stubbe N, Milch R, Parker R, Rasmussen KM, Root M, Graham S, et al. Breast cancer and dietary and plasma concentrations of carotenoids and vitamin A. Am J Clin Nutr. 1990 Nov;52(5):909–915. [PubMed]
  • Koga M, Sutherland RL. Retinoic acid acts synergistically with 1,25-dihydroxyvitamin D3 or antioestrogen to inhibit T-47D human breast cancer cell proliferation. J Steroid Biochem Mol Biol. 1991 Oct;39(4A):455–460. [PubMed]

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