Retinoic acid nuclear receptor β inhibits breast carcinoma anchorage independent growth
Xiao-Su Li
Department of Medicine, Division of Oncology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
University of Maryland, School of Medicine and the Veterans Administration Medical Center, Baltimore, Maryland 21201
Search for more papers by this authorZhi-Ming Shao
Department of Medicine, Division of Oncology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
University of Maryland, School of Medicine and the Veterans Administration Medical Center, Baltimore, Maryland 21201
Search for more papers by this authorM. Saeed Sheikh
Department of Medicine, Division of Oncology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
University of Maryland, School of Medicine and the Veterans Administration Medical Center, Baltimore, Maryland 21201
Search for more papers by this authorJulie L. Eiseman
Departments of Pathology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
Search for more papers by this authorDorothy Sentz
Departments of Pathology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
Search for more papers by this authorAnton M. Jetten
Cell Biology Section, Laboratory of Pulmonary Pathobiology, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709
Search for more papers by this authorJian-Chyi Chen
Department of Medicine, Division of Oncology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
Search for more papers by this authorMarcia I. Dawson
Life Sciences Division, SRI International, Menlo Park, California 94025
Search for more papers by this authorSeena Aisner
Departments of Pathology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
Search for more papers by this authorArun K. Rishi
Department of Medicine, Division of Oncology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
Search for more papers by this authorPeter Gutierrez
Departments of Biochemistry, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
Search for more papers by this authorLauren Schnapper
Departments of Sugery, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
Search for more papers by this authorCorresponding Author
Joseph A. Fontana
Department of Medicine, Division of Oncology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
University of Maryland, School of Medicine and the Veterans Administration Medical Center, Baltimore, Maryland 21201
University of Maryland at Baltimore, Cancer Center, Room S9 D05, 22 S. Greene Street, Baltimore, Maryland 21201Search for more papers by this authorXiao-Su Li
Department of Medicine, Division of Oncology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
University of Maryland, School of Medicine and the Veterans Administration Medical Center, Baltimore, Maryland 21201
Search for more papers by this authorZhi-Ming Shao
Department of Medicine, Division of Oncology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
University of Maryland, School of Medicine and the Veterans Administration Medical Center, Baltimore, Maryland 21201
Search for more papers by this authorM. Saeed Sheikh
Department of Medicine, Division of Oncology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
University of Maryland, School of Medicine and the Veterans Administration Medical Center, Baltimore, Maryland 21201
Search for more papers by this authorJulie L. Eiseman
Departments of Pathology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
Search for more papers by this authorDorothy Sentz
Departments of Pathology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
Search for more papers by this authorAnton M. Jetten
Cell Biology Section, Laboratory of Pulmonary Pathobiology, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709
Search for more papers by this authorJian-Chyi Chen
Department of Medicine, Division of Oncology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
Search for more papers by this authorMarcia I. Dawson
Life Sciences Division, SRI International, Menlo Park, California 94025
Search for more papers by this authorSeena Aisner
Departments of Pathology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
Search for more papers by this authorArun K. Rishi
Department of Medicine, Division of Oncology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
Search for more papers by this authorPeter Gutierrez
Departments of Biochemistry, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
Search for more papers by this authorLauren Schnapper
Departments of Sugery, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
Search for more papers by this authorCorresponding Author
Joseph A. Fontana
Department of Medicine, Division of Oncology, Baltimore, Maryland 21201
Departments of Cancer Center, Baltimore, Maryland 21201
University of Maryland, School of Medicine and the Veterans Administration Medical Center, Baltimore, Maryland 21201
University of Maryland at Baltimore, Cancer Center, Room S9 D05, 22 S. Greene Street, Baltimore, Maryland 21201Search for more papers by this authorAbstract
Retinoids modulate cellular proliferation and mediate gene function through a series of nuclear receptors. The retinoic acid nuclear receptor β (RARβ) plays an important role in the differentiation of a number of cell types. We now demonstrate that RARβ expresion is confined to normal mammary tissue and is not expressed in either immortalized normal or malignant cell lines. Treatment of RARβ-transfected MDA-MB-231 cells with 1 μM all-trans-retinoic acid (RA) significantly inhibited monolayer growth of the cells which express recombinant RARβ. RARβ-expressing MDA-MB-231 cells formed significantly smaller and fewer colonies soft agar than the mock-transfected cells. Addition of 1 μM RA stimulated colony size and number in the RARβ-transfected MDA-MB-231 cells. In contast to the RARβ-expressing cells, colony formation by the RARβ-expressing cells was similar to the mock-transfected controls and the addition of 1 μM RA to the RARα-transfected cells inhibited colony formation. While demonstrating decreased colony formation in agar, RARβ-expressing MDA-MB-231 cells failed to exhibit decreased growth in SCID mice. Our results show that RARβ functions as a negative regulator of growth in breast epithelial cells. In addition, the growth of these cells is differentially regulated by RARα and RARβ which is most likely the result to the modulation of different genes. © 1995 Wiley-Liss Inc.
Literature Cited
- Allenby, G., Janocha, R., Kazmer, S., Speck, J., Gripp, J. E., and Levin, A. A. (1994) Binding of 9-cis-retinoic acid and all-trans-retinoic acid to retinoic acid receptors alpha, beta, and gamma. Retinoic acid receptor gamma binds all-trans-retinoic acid preferentially over 9-cis-retinoic acid. J. Biol. Chem., 269: 16689–16695.
- Benbrook, D., Lernhardt, E., and Pfahl, M. (1988) A new retinoic acid receptor identified from a hepatocellular carcinoma. Nature (Lond.), 333: 669–672.
- Brand, N., Petkovich, M., Krust, A., Chambon, P., de Thé, H., Marchio, A., Tiollais, P., and Dejean, A. (1988) Identification of a second human retionic acid receptor. Nature (Lond.), 332: 850–853.
- Dawson, M. I., Hobbs, P. D., Derdzinski, K. A., Chao, W.-R., Frenking, G., Loew, G. H., Jetten, A. M., Napoli, J. L., Williams, J. B., Sani, B. P., and Wille, J. (1989) Effect of structural modifications in the C7-C11 region of the retinoid skeleton on biological activity in a series of aromatic retinoids. J. Med. Chem., 32: 1504–1517.
- de Thé, H., Marchio, A., Tiollais, P., and Dejean, A. (1989) Differential expression and ligand regulation of the retinoic acid receptor α and β genes. EMBO J., 8: 429–433.
- Fanjul, A., Dawson, M. I., Hobbs, P. R., Jong, L., Cameron, J. F., Harlev, E., Graupner, G., Lu, X.-P., and Pfahl, M. (1994) A new class of retinoids with selective inhibition of AP-1 inhibits proliferation. Nature, 372: 107–111.8.
- Fontana, J. A. (1987) Interaction of retinoids and tamoxifen on the inhibition of human mammary carcinoma cell proliferation. Exp. Cell Biol., 55: 136–144.
- Fontana, J. A., Miksis, G., Miranda, D. M., and Durham, J. (1987) Inhibition of human mammary carcinoma cell proliferation by retinoids and intracellular cAMP-elevating compounds. J. Natl. Cancer Inst., 78: 1107–1112.
- Fontana, J. A., Nervi, C., Shao, Z.-M., and Jetten, A. M. (1992) Retinoid antagonism of estrogen-responsive TGF-α and pS2 expression in breast carcinoma cells. Cancer Res., 52: 3838–3945.
- Gaetano, C., Matsumoto, K., and Thiele, C. J. (1991) Retinoic acid negatively regulates p34cdc2 expression during human neuroblastoma differentiation. Cell Growth Differ., 2: 487–493.
- Gebert, J. F., Moghal, N., Frangioni, J. V., Sugarbaker, D. J., and Neel, B. G. (1991) High frequency of retinoic acid receptor β abnormalities in human lung cancer. Oncogene, 6: 1859–1868.
- Giguere, V. (1994) Retinoic acid receptors and cellular retinoid binding proteins: Complex interplay in retinoid signaling. Endoc. Rev., 15: 61–79.
- Giguere, V., Ong, E. S., Segui, P., and Evans, R. M. (1987) Identification of a receptor for the morphogen retinoic acid. Nature (Lond.), 330: 624–629.
- Gudas, L. J. (1992) Retinoids, retinoic-responsive genes, cell differentiation, and cancer. Cell Growth Differ., 3: 655–662.
- Gudas, L. J. (1994) Retinoids and vertebrate development. J. Biol. Chem., 269: 15399–15402.
- Houle, B., Leduc, F., and Bradley, W. E. C. (1991) Implication of RARβ in epidermoid (squamous) lung cancer. Genes Chrom. Cancer, 3: 358–366.
- Houle, B., Rochette-Egly, C., and Bradley, W. E. C. (1993) Tumor suppressive effect of the retinoic acid receptor β in human epidermoid lung cancer cells. Proc. Natl. Acad. Sci. U.S.A., 90: 985–989.
- Houle, B., Pelletier, M., Wu, J., Goodyer, C., and Bradley, W. E. C. (1994) Fetal isoform of human retinoic acid receptor β expressed in small cell lung cancer cell lines. Cancer Res., 54: 365–369.
- Hu, L., and Gudas, L. J. (1990) Cyclic AMP analogs and retinoic acid influence the expression of retinoic acid receptors alpha, beta, and gamma mRNAs in F9 teratocarcinoma cells. Mol. Cell. Biol., 10: 391–396.
- Hu, L., Crowe, D. L., Rheinwald, J. G., Chambon, P., and Gudas, L. J. (1991) Abnormal expression of retinoic acid receptors and keratin 19 by human oral and epidermal squamous cell carcinoma cell lines. Cancer Res., 51: 3972–3981.
- Jetten, A. M., Brody, A. R., Deas, M. A., Hook, G. E. R., Rearick, J. L., and Thacher, S. M. (1987) Retinoic acid and substratum regulate the differentiation of rabbit tracheal epithelial cells into squamous and secretory phenotype. Morphological and biochemical characterization. Lab. Invest., 56: 654–664.
- Kagechika, H., Kawachi, E., Hashimoto, Y., Himi, T., and Shudo, K. (1988) Retinobenzoic acids. 1. Structure-activity relationships of aromatic amides with retinoidal activity. J. Med. Chem., 31: 2182–2192.
- Koga, M., and Sutherland, R. L. (1991) Retinoic acid acts synergistically with 1,25-dihydro vitamin D3 or antiestrogen to inhibit T-47D human breast cancer cell proliferation. J. Steroid Biochem. Mol. Biol., 39: 455–460.
- Krust, A., Kastner, P., Petkovich, M., Zelent, A., and Chambon, P., (1989) A third human retinoic acid receptor, hRARγ, Proc. Natl. Acad. Sci. U.S.A., 86: 5310–5314.
- Lehman, J. M., Dawson, M. I., Hobbs, P. O., Husmann, M., and Pfahl, M. (1991) Identification of retinoids with nuclear receptor subtype-selective activities. Cancer Res., 51: 4807–4809.
- Leid, M., Kastner, P., Lyons, R., Nakshatri, H., Saunders, M., Zacharewski, T., Chen, J.-Y., Staub, A., Garnier, J.-M., Mader, S., and Chambon, P. (1992) Purification, cloning and RXR identity of the HeLa cell factor with which RAR or TR heterodimerizes to bind target sequences efficiently. Cell, 68: 377–395.
- Levin, A. A., Sturzenbecker, L. M., Kazmer, S., Bosakowski, T., Huselton, C., Allenby, G., Speck, J., Kratzeisen, C., Rosenberger, M., Lovey, A., and Grippo, J. F. (1992) 9-cis-retinoic acid stereoisomer binds and activates the nuclear receptor RXR alpha. Nature, 355: 359–361.
- Li, E., Sucov, H. M., Lee, K.-F., Evans, R. M., and Jaenisch, R. (1993) Normal development and growth of mice carrying a targeted disruption of the alpha 1 retinoic acid receptor gene. Proc. Natl. Acad. Sci. U.S.A., 90: 1590–1594.
- Li, X.-S., Chen, J.-C., Sheikh, M. S., Shao, Z.-M., and Fontana, J. A. (1994) Retinoic acid inhibition of insulin-like growth factor I stimulation of c-fos mRNA levels in a breast carcinoma cell line. Exp. Cell Res., 211: 68–83.
- Lohnes, D., Kastner, P., Dierich, A., Mark, M., LeMeur, M., and Chambon, P. (1993) Function of retinoic acid receptor gamma in the mouse. Cell, 73: 643–658.
- Lufkin, T., Lohnes, D., Mark, M., Dierich, A., Gorry, P., Gaub, M.-P., LeMeur, M., and Chambon, P. (1993) High postnatal lethality and testis degeneration in retinoic acid receptor alpha mutant mice. Proc. Natl. Acad. Sci. U.S.A., 90: 7225–7229.
- Mangelsdorf, D. J., Borgmeyer, U., Heyman, R. A., Zhou, J. Y., Ong, E. S., Oro, A. E., and Kakizuka, A. (1992) Characterization of three RXR genes that mediate the action of 9-cis retinoic acid. Genes Dev., 5: 329–344.
- Mattei, M.-G., de Thé, H., Mattei, J. F., Marchio, A., Tiollais, P., and DeJean, A. (1988) Assignment of the human hap retinoic acid receptor RAR beta gene to the p24 band of chromosome 3. Human Genet., 80: 189–190.
- Moasser, M. M., DeBlasso, A., and Dmitrovsky, E. (1994) Response and resistance to retinoic acid are mediated through the retinoic acid nuclear receptor gamma in human teratocarcinomas. Oncogene, 9: 833–840.
- Nagpal, S., Saunders, M., Kastner, P., Durand, B., Nakshatri, H., and Chambon, P. (1992) Promoter context- and response element-dependent specificity of the transcriptional activation and modulating functions of retinoic acid receptors. Cell, 70: 1007–1019.
- Negrini, M., Sabbioni, S., Haldor, S., Possati, L., Castagnoli, A., Corallini, A., Barbanti-Brodano, G., and Croce, C. M. (1994) Tumor and growth suppression of breast cancer cells by chromosome 17-associated functions. Cancer Res., 54: 1818–1824.
- Nervi, C., Vollberg, T. M., George, M. D., Zelent, A., Chambon, P., and Jetten, A. M. (1991) Expression of nuclear retinoic acid receptors in normal tracheobronchial cells and in lung carcinoma cells. Exp. Cell Res., 195: 163–170.
- Ng, S-Y., Gunning, P., Eddy, R., Ponte, P., Leavitt, J., Shows, T., and Kedes, L. (1985) Evolution of the functional human β-actin gene and its multi-pseudogene family: Conservation of noncoding regions and chromosomal dispersion of pseudogenes. Mol. Cell. Biol. 5: 2720–2732.
- Nicholson, R. C., Mader, S., Nagpal, S., Leid, M., Rochette-Egly, C., and Chambon, P. (1990) Negative regulation of the RAT Stromelysin gene promoter by retinoic acid is mediated by an AP-1 binding site. EMBO J., 9: 4443–4454.
- Petkovich, M., Brand, N. J., Krust, A., and Chambon, P. (1987) A human retinoic acid receptor which belongs to the family of nuclear receptors. Nature (Lond.), 330: 444–450.
- Pierce, J. H., Arnstein, P., DiMarco, E., Artrip, J., Kraus, M. H., Lonardo, F., DiFiore, P. P., and Aaronson, S. A. (1991) Oncogenic potential of erbB-2 in human mammary epithelial cells. Oncogene, 6: 1189–1194.
- Reichel, R. R. (1992) Regulation of E2F/cyclin A-containing complex upon retinoic acid-induced differentiation of teratocarcinoma cells. Gene Exp., 2: 259–271.
- Roman, S. D., Clarke, C. L., Hall, R. E., Alexander, I. E., and Sutherland, R. L. (1992) Expression and regulation of retinoic acid receptors in human breast cancer cells. Cancer Res., 52: 2236–2242.
- Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Expression of cloned genes in cultured mammalian cells. In: Molecular Cloning. A Laboratory Manual, 2nd ed. N. Ford and C. Nolan, eds. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, pp. 16.33–16.36.
- Sawyers, C. L., McLaughlin, J., Goga, A., Havlik, M., and Witte, O. (1994) The nuclear tyrosine kinase c-abl negatively regulates cell growth. Cell, 77: 121–131.
- Schiff, L. J., Okamura, W. H., Dawson, M. I., and Hobbs, P. D. (1990) Chemistry and Biology of Synthetic Retinoids. M. I. Dawson and W. H. Okamura, eds. CRC Press, Boca Raton, FL, p. 321.
- Schüle, R., Rangarojan, P., Yang, N., Kliewer, S., Ransone, L. J., Bolado, J., Verda, I. M., and Evans, R. M. (1991) Retinoic acid is a negative regulator of AP-1 responsive genes. Proc. Natl. Acad. Sci. U.S.A., 88: 6092–6096.
- Shao, Z.-M., Sheikh, M. S., Chen, J.-C., Kute, T., Aisner, S., Schnaper, L., and Fontana, J. A. (1994) Expression of the retinoic acid nuclear receptors (RARs) and retinoid X receptor (RXR) in estrogen receptor positive and negative breast cancer. Int. J. Oncol., 4: 849–853.
- Sheikh, M. S., Shao, Z-M, Chen, J-C., Hussain, A., Jetten, A. M., and Fontana, J. A. (1993) Estrogen receptor-negative breast cancer cells transfected with the estrogen receptor exhibit increased RARα gene expression and sensitivity to growth inhibition by retinoic acid. J. Cell Biochem., 53: 394–404.
- Sheikh, M. S., Shao, Z. M., Li, X., Dawson, M. I., Jetten, A. M., Conley, B. A., Garcia, M., Rochefort, H., and Fontana, J. A. (1994) Retinoid-resistant estrogen receptor-negative human breast carcinoma cells transfected with retinoic acid receptor-α acquire sensitivity to growth inhibition by retinoids. J. Biol. Chem., 269: 21440–21447.
- Shin, S., Freedman, V. H., Risser, R., and Pollack, R. (1975) Tumor-genicity of virus-transformed cells in nude mice is correlated specifically with anchorage-independent growth in vitro. Proc. Natl. Acad. Sci. U.S.A., 72: 4435–4439.
- Soule, H. D., Maloney, T. M., Wolman, S. R., Peterson, W. D., Jr., Brenz, R., McGrath, C. M., Russo, J., Pauley, R. J., Jones, R. F., and Brooks, S. C. (1990) Isolation and characterization of a spontaneously immortalized human breast epithelial cell line, MCF-10. Cancer Res., 50: 6075–6086.
- Southern, P. J., and Berg, P. (1982) Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. Mol. Appl. Genet., 1: 327–341.
- Stampfer, M. R., and Bartley, J. C. (1985) Induction of transformation and continuous cell lines from normal human mammary epithelial cells after exposure to benzopyrene. Proc. Natl. Acad. Sci. U.S.A., 82: 2394–2398.
- Swisshelm, K., Ryan, K., Lee, X., Tsou, H. C., Peacocue, M., and Sager, R. (1994) Down-regulation of retinoic acid receptor β in mammary carcinoma cell lines and its up-regulation in senescing normal mammary epithelial cells. Cell Growth Differ., 5: 133–141.
- Zelent, A., Mendelson, C., Kastner, P., Krust, A., Garnier, J. M., Ruffenach, F., Leroy, P., and Chambon, P. (1991) Differentially expressed isoforms of the mouse retinoic acid receptor beta generated by usage of two promoters and alternative splicing. EMBO J., 10: 71–81.
- Zhang, X-K, Li, Y., Lee, M.-O., and Pfahl, M. (1994) A specific defect in the retinoic acid response associated with human lung cancer cell lines. Cancer Res., 54: 5663–5469.