Abstract
Retinoids constitute a very promising class of agents for the chemoprevention or treatment of breast cancer. These retinoids exert their biological activity through two distinct classes of retinoic acid (RA) receptors (R), the RAR isotypes (α, β, and γ) and the three RXR isotypes (α, β, and γ) and their numerous isoforms which bind as RXR/RAR heterodimers to the polymorphic cis-acting response elements of RA target genes. With respect to these numerous receptor sub-types, the retinoid-induced effects at the biological level include marked modifications with respect to both cell proliferation and cell death (apoptosis), and also in the induction of differentiation processes. The present study aims to characterize the effect which four retinoids (TTNPB, 9-cis-RA, LGD 1069, 4-HPR) with distinct RAR/RXR binding properties induced on various in vitro and in vivo mouse and human breast cancer models. The experiments with the retinoids were carried out in comparison with the anti-estrogen tamoxifen and the anti-progestagen RU-486 compounds. The results show that the 6 compounds under study were markedly more efficient in terms of growth inhibition in the human T-47D cell line when maintained under anchorage-independent culture conditions than when maintained under anchorage-dependent ones. While RU-486 exhibited a weak statistically significant (p < 0.05) influence on the growth of the T-47D stem cells, tamoxifen had a marked inhibitory influence on the growth of these cells. Of the four retinoids, 4-HPR was the least effective since the lowest doses tested (1 and 0.1 nM) exhibited no statistically (p > 0.05) significant influence on the growth of the stem cells. The most efficient retinoid was TTNPB. It was only at the highest dose (10 μM) that tamoxifen and RU-486 showed a weak inhibitory influence on the growth of the T-47D non-stem cells while all 4 retinoids exerted a significant inhibitory influence on the growth of these non-stem cells, with 4-HPR being the most efficient (P < 0.001) at the highest dose, but ineffective (P > 0.05) at the lowest. Tamoxifen and TTNPB were tested in vivo on hormone-senstive (HS) and hormone-insensitive (HI) strains of the MXT murine mammary carcin oma. While TTNPB appeared to be equally efficient in terms of growth inhibition in both MXT-HS and MXT-HI models, tamoxifen had only a marginal inhibitory influence on the growth of the MXT-HI strain but did inhibit growth in the case of the MXT-HS one. TTNPB was markedly more efficient than tamoxifen in terms of both inhibiting the cell proliferation level (measured by means of computer-assisted microscopy applied to Feulgen-stained nuclei, a method which enables the percentage of cells in the S phase of the cell cycle to be determined) and triggering cell death (measured by means of the determination of the transglutaminase activity) in both the MXT-HI and MXT-HS models. The very significant TTNPB-induced inhibition of the macroscopic MXT-HS growth rate relates to the triggering of cell death (apoptosis) rather than to an inhibition of cell proliferation. All these results clearly indicate that retinoids are very efficient agents against breast cancer, at least as efficient as tamoxifen.
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References
Fisher B, Osborne CK, Margolese R, Bloomer W: Neoplasms of the breast. In: Holland JF, Frei E III, Bast RC Jr, Kufe DW, Morton DL, Weichselbaum RR (eds) Cancer Medicine. Lea & Febiger, Philadelphia-London, Volume 1, 1993, pp T706–T774
Beatson GT: On the treatment of inoperable cases of carcinoma of the mamma. Suggestion for a new method of treatment with illustrative cases. Lancet 2: 104–107, 1896
Paridaens R, Leclercq G, Piccart M, Kiss R, Mattheiem W, Heuson JC: Comments on the treatment of breast cancer. In: Bulbrook RD (ed) Hormones of Cancer: 90 Years after Beatson. Cancer Surveys 5, 1986, pp 447–461
Costa A, Sacchini V, Decensi A: Retinoids and tamoxifen in breast cancer chemoprevention. Int J Clin Lab Res 23: 53–55, 1993
Anzano MA, Byers SW, Smith JM, Peer CM, 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 54: 4614–4617, 1994
Gottardis MM, Bischoff ED, Shirley MA, Wagoner MA, Lamph WW, Heyman RA. Chemoprevention of mammary carcinoma by LGD1069 (Targretin): an RXR-selective ligand. Cancer Res 56: 5566–5570, 1996
Cassano E, Coopmans de Yoldi G, Ferranti C, Costa A, Mascotti G, De Palo G, Veronesi U: Mammographic patterns in breast cancer chemoprevention with Fenretinide (4-HPR). Eur J Cancer 29A: 2161–2163, 1993
Gottardis MM, Lamph WW, Shalinsky DR, Wellstein A, Heyman RA: The efficacy of 9-cis-retinoic acid in experimental models of cancer. Breast Cancer Res Treat 38: 85–96, 1996
Dawson MI, Chao WR, Pine P, Jong L, Hobbs PD, Rudd CK, Quick TC, Niles RM, Zhang XK, Lombardo A, Ely KR, Shroot B, Fontana JA: Correlation of retinoic binding affinity to retinoic acid receptor α with retinoid inhibition of growth of estrogen receptor-positive MCF-7 mammary carcinoma cells. Cancer Res 55: 4446–4451, 1995
Shao ZM, Dawson MI, Li XS, Rishi AK, Sheikh MS, Han QX, Ordonez JV, Shroot B, Fontana JA: p53 independent G0/G1 arrest and apoptosis induced by a novel retinoid in human breast cancer cells. Oncogene 11: 493–504, 1995
James SY, Mackay AG, Colston KW: Vitamin D derivatives in combination with 9-cis-retinoic acid promote active cell death in breast cancer cells. J Mol Endocrinol 14: 391–394, 1995
Fanjul AN, Bouterfa H, Dawson M, Pfahl M: Potential role for retinoic acid receptor γ in the inhibition of breast cancer cells by selective retinoids and interferons. Cancer Res 56: 1571–1577, 1996
Liu Y, Lee MO, Wang HG, Li Y, Hashimoto Y, Klaus M, Reed JC, Zhang X: Retinoic acid receptor β mediates the growth-inhibitory effect of retinoic acid by promoting apoptosis in human breast cancer cells. Mol Cell Biol 16: 1138–1149, 1996
Lotan R: Retinoids in cancer chemoprevention. FASEB J 10: 1031–1039, 1996
Chambon P: A decade of molecular biology of retinoic acid receptors. FASEB J 10: 940–954, 1996
Urbach J, Rando RR: Isomerization of all-trans-retinoic acid to 9-cis-retinoic acid. Biochem J 299: 459–465, 1994
Boehm MF, Zhang L, Badea BA, White SK, Mais DE, Berger E, Suto CM, Goldman ME, Heyman RA: Synthesis and structure-activity relationship of novel retinoid X receptor selective retinoids. J Med Chem 37: 2930–2941, 1994
Beard RL, Gil DW, Marler DK, Henry E, Colon DF, Gillett SJ, Arefieg T, Breen TS, Krauss H, Davies PJA, Chandraratna RAS: Structural basis for the differential RXR and RAR activity of stilbene retinoid analogs. Biorg Med Chem Lett 4: 1447–1452, 1994
Pellegrini R, Mariotti A, Tagliabue E, Bressau R, Bunone G, Coradini D, Della Valle G, Formelli F, Cleris L, Radice P, Pierotti MA, Colnghi MI, Ménard S: Modulation of markers associated with tumor aggressiveness in human breast cancer cell lines by N-(4-hydroxyphenyl) retinamide. Cell Growth Differ 6: 863–869, 1995
Kizaki M, Dawson MI, Heyman R, Elster E, Morosetti R, Pakkala S, Chen DL, Ueno H, Chao W, Morikawa M, Ikeda Y, Heber D, Pfahl M, Koeffler HP: Effects of novel retinoid X receptor-selective ligands on myeloid leukemia differentiation and proliferation in vitro. Blood 87: 1977–1984, 1996
Beck CA, Zhang Y, Weigel NL, Edwards DP: Two types of anti-progestins have distinct effects on site-specific phosphorylation of human progesterone receptor. J Biol Chem 271: 1209–1217, 1996
Watson CS, Medina D, Clark JH: Estrogen receptor characterization in a transplantable mouse mammary tumor. Cancer Res 37: 3344–3348, 1977
Danguy A, Kiss R, Leclercq G, Heuson JC, Pasteels JL: Morphology of MXT mouse mammary tumors. Correlation with growth characteristics and hormone sensitivity. Eur J Cancer Clin Oncol 22: 69–76, 1986
Kiss R, de Launoit Y, Danguy A, Paridaens R, Pasteels JL: Influence of pituitary grafts or prolactin administrations on the hormone sensitivity of ovarian hormone-independent mouse mammary MXT tumors. Cancer Res 49: 2945–2951, 1989
Briand P: Hormone-dependent mammary tumors in mice and rats as a model for human breast cancer (review). Anti-cancer Res 3: 273–282, 1983
Fingert HJ, Campisi J, Pardee AB: Cancer biology. Cell proliferation and differentiation. In: Holland JF, Frei E III, Bast RC Jr, Kufe DW, Morton DL, Weichselbaum RR (eds) Cancer Medicine. Lea & Febiger, Philadelphia-London, Volume 1, 1993, pp 1–14
Piacentini M, Fesus L, Farrace MG, Ghibelli L, Piredda L, Melino G: The expression of ‘tissue’ transglutaminase in two human cancer cell lines is related with the programmed cell death (apoptosis). Eur J Cell Biol 54: 246–254, 1991
Horn V, Minucci S, Ogryzko VV, Adamson ED, Howard BH, Levein AA, Ozato K: RAR and RXR selective ligands cooperatively induce apoptosis and neuronal differentiation in P19 embryonal carcinoma cells. FASEB J 10: 1071–1077, 1996
Salmon I, Kiss R: Relationship between proliferative activity and ploidy level in a series of 530 human brain tumors including astrocytomas, meningiomas, schwannomas and metastases. Human Pathol 24: 329–335, 1993
Freake HC, Marcocci C, Iwasaki J, MacIntyre I: 1,25-dihydroxyvitamin D3 specifically binds to a human breast cancer cell line (T47D) and stimulates growth. Biochem Biophys Res Commun 101(4): 1131–1138, 1981
Camby I, Salmon I, Danguy A, Pasteels JL, Brotchi J, Martinez J, Kiss R: Influence of gastrin on human astrocytic tumor cell proliferation. J Natl Cancer Inst 88: 594–600, 1996
Janssen T, Darro F, Petein M, Raviv G, Pasteels JL, Kiss R, Schulman C: In vitro characterization of prolactin-induced effects on proliferation in the neoplastic LNCaP, DU145 and PC3 models of the human prostate. Cancer 77: 144–149, 1996
Mosman T: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxic assay. J Immunol Meth 65: 55–63, 1983
Vistica DT, Skehan P, Scudiero D, Monks A, Pittman A, Boyd MR: Tetrazolium-based assays for cellular viability: a critical examination of selected parameters affecting formazan production. Cancer Res 51: 2515–2525, 1991
Rubinstein LV, Shoemaker RH, Paull KD, Simon RM, Tosini S, Skehan P, Scudiero DA, Monks A, Boyd MR: Comparison of in vitro anticancer-drug-screening data generated with tetrazolium assay versus a protein assay against a diverse panel of human tumor cell lines. J Natl Cancer Inst 82: 1113–1118, 1990
Kiss R, Salmon I, Camby I, Gras S, Pasteels JL: Characterization of factors in routine laboratory protocols that significantly influence the Feulgen reaction. J Histochem Cytochem 41: 935–945, 1993
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275, 1951
Darzynkiewicz Z, Bruno S, Del Bino G, Gorczyca W, Hotz MA, Lassota P, Traganos F: Features of apoptotic cells measured by flow cytometry. Cytometry 13: 795–808, 1992
Ponzoni M, Bocca P, Chiesa V, Decensi A, Pistoia V, Raffaghello L, Rozzo C, Montaldo PG: Differential effects of N-(4-hydroxyphenyl)retinamide and retinoic acid on neuroblastoma cells: apoptosis versus differentiation. Cancer Res 55: 853–861, 1995
Sporn MB, Newton DL: Chemoprevention of cancer with retinoids. Fed Proc 38: 2528–2534, 1979
Jing Y, Zhang J, Waxman S, Mira Y, Lopez R: Upregulation of cytokeratins 8 and 18 in human breast cancer T-47D cells is retinoid-specific and retinoic acid receptor-dependent. Differentiation 60: 109–117, 1996
Van Der Leede BM, Geertzema J, Vroom TM, Decimo D, Lutz Y, Van Der Saag PT, Van Der Burg B: Immunohistochemical analysis of retinoic acid receptor-alpha in human breast tumors: retinoic acid receptor-alpha expression correlates with proliferative activity. Am J Pathol 148: 1905–1914, 1996
Wang TT, Phang JM: Effect of N-(4-hydroxyphenyl)retinamide on apoptosis in human breast cancer cells. Cancer Lett 107: 65–71, 1996
Fanjul AN, Delia D, Pierotti MA, Rideout D, Yu JQ, Pfahl M: 4-hydroxyphenyl retinamide is a highly selective activator of retinoid receptors. J Biol Chem 271(37): 22441–22446, 1996
Kazmi SM, Plante RK, Visconti V, Lau CY: Comparison of N-(4-hydroxyphenyl)retinamide and all-trans-retinoic acid in the regulation of retinoid receptor-mediated gene expression in human breast cancer cell lines. Cancer Res 56: 1056–1062, 1996
Oridate N, Suzuki S, Higuchi M, Mitchell MF, Hong WK, Lotan R: Involvement of reactive oxygen species in N-(4-hydroxyphenyl)retinamide-induced apoptosis in cervical carcinoma cells. J Natl Cancer Inst 89(16): 1191–1198, 1997
Standeven AM, Johnson AT, Escobar M, Chandraratna RA: Specific antagonist of retinoid toxicity in mice. Toxicol Appl Pharmacol 138: 169–175, 1996
Paridaens RJ, Danguy AJ, Leclercq G, Kiss R, Heuson JC: Effect of castration and 17 β-estradiol pulse on cell proliferation in the uterus and the MXT mouse mammary tumor. J Natl Cancer Inst 74: 1239–1246, 1985
Harris J, Morrow M, Norton L: Cancer of the breast (Chapter 36). Malignant tumors of the breast (Section 2). In: De Vita VT Jr, Hellman S, Rosenberg SA (eds) Cancer: Principles and Practice of Oncology. Fifth Edition. JB Lippincott Co., Philadelphia, 1997, pp 1557–1616
Pollack IF, Kawccki S: The efficacy of tamoxifen as an anti-proliferative agent in vitro for benign and malignant pediatric glial tumors. Pediatric Surg 22: 281–288, 1995
Couldwell WT, Hinton DR, He S, Chen TC, Sebat I, Weiss MH, Law RE: Protein kinase C inhibitors induce apoptosis in human malignant glioma cell lines. FEBS Lett 345: 43–46, 1994
Berry J, Green BJ, Matheson DS: Modulation of natural killer cell activity by tamoxifen in stage I post-menopausal breast cancer. Eur J Cancer Clin Oncol 23: 517–520, 1987
Gottardis MM, Wagner RJ, Borden EC, Jordan VC: Differential ability of antiestrogens to stimulate breast cancer cell (MCF-7) growth in vitro and in vivo. Cancer Res 49: 4765–4769, 1989
Szende B, Schally AV, Comaru-Schally AM, Redding TW, Srkalovic G, Groot K, Lapis K, Timar J, Neill J, Mulchahey J: Cellular and molecular aspects of apoptosis in experimental tumors of animals treated with analogs of LHRH and somatostatin. In: Tomei LD, Cope FO (eds) Apoptosis: The Molecular Basis of Cell Death. Cold Spring Laboratory Press, 1991, pp 139–155
Allegretto EA, McClurg MR, Lazarchik SB, Clemm DL, Kerner SA, Elgort MG, Boehm MF, White SK, Pike JW, Heyman RA: Transactivation properties of retinoic acid and retinoid X receptors in mammalian cells and yeast. J Biol Chem 258: 26625–26633, 1993
Pignatello MA, Kauffman FC, Levin AA: Multiple factors contribute to the toxicity of the aromatic retinoid, TTNPB (Ro 13-7410): binding affinities and disposition. Toxicol Appl Pharmacol 142: 319–327, 1997
Gottardis MM, Bischoff ED, Shirley MA, Wagoner MA, Lamph WW, Heyman RA: Chemoprevention of mammary carcinoma LGD1069 (targretin): an RXR-selective ligand. Cancer Res 56: 5566–5570, 1996
Sheikh MS, Zhao Z-M, Li X-S, Ordonez JV, Conley BA, Wu S, Dawson MJ, Han Q-X, Chao W-R, Quick T, Niles RM, Fontana JA: N-(4-hydroxyphenyl)retinamide (4-HPR)-mediated biological actions involve retinoid receptor-independent pathways in human breast carcinoma. Carcinogenesis 16: 2477–2486, 1995
Viljoen TC, van Aswegen CH, du Plessis DJ: Binding of estradiol to whole prostatic DU-145 cells in the presence and absence of tamoxifen and acetylsalicylic acid. Prostate 27: 160–165, 1995
Terakawa N, Shimizu I, Tanizawa O, Matsumoto K: RU486, a progestin antagonist, binds to progesterone receptors in a human endometrial cancer cell line and reverses the growth inhibition by progestins. J Steroid Biochem 31: 161–166, 1988
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Darro, F., Cahen, P., Vianna, A. et al. Growth inhibition of human in vitro and mouse in vitro and in vivo mammary tumor models by retinoids in comparison with tamoxifen and the RU-486 anti-progestagen. Breast Cancer Res Treat 51, 39–55 (1998). https://doi.org/10.1023/A:1006098124087
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DOI: https://doi.org/10.1023/A:1006098124087