Preferences help
enabled [disable] Abstract
Number of results
2004 | 51 | 3 | 733-745
Article title

Can transforming growth factor-β1 and retinoids modify the activity of estradiol and antiestrogens in MCF-7 breast cancer cells?

Title variants
Languages of publication
Retinoic acid and transforming growth factor-β (TGF-β) affect differentiation, proliferation and carcinogenesis of epithelial cells. The effect of both compounds on the proliferation of cells of the hormone sensitive human breast cancer cell line (ER+) MCF-7 was assessed in the presence of estradiol and tamoxifen. The assay was based on [3H]thymidine incorporation and the proliferative activity of PCNA- and Ki 67-positive cells. The apoptotic index and expression of the Bcl-2 and p53 antigens in MCF-7 cells were also determined. Exogenous TGF-β1 added to the cell culture showed antiproliferative activity within the concentration range of 0.003-30 ng/ml. Irrespective of TGF-β1 concentrations, a marked reduction in the stimulatory action of estradiol (10-9 and 10-8 M) was observed whereas in combination with tamoxifen (10-7 and 10-6 M) only 30 ng/ml TGF-β1 caused a statistically significant reduction to aproximately 30% of the proliferative cells. In further experiments we examined the effect of exposure of breast cancer cells to retinoids in combination with TGF-β1. The incorporation of [3H]thymidine into MCF-7 cells was inhibited to 52 ± 19% (control =100%) by 3 ng/ml TGF-β1, and this dose was used throughout. It was found that addition of TGF-β1 and isotretinoin to the culture did not decrease proliferation, while TGF-β1 and tretinoin at low concentrations (3 × 10-8 and 3 × 10-7 M) reduced the percentage of proliferating cells by aproximately 30% (67±8% and 67±5%, P <0.05 compared to values in the tretinoin group). Both retinoids also led to a statistically significant decrease in the stimulatory effect of 10-9 M estradiol, attenuated by TGF-β1. In addition, the retinoids in combination with TGF-β1 and tamoxifen (10-6 M) caused a further reduction in the percentage of proliferating cells. Immunocytochemical analysis showed that all the examined compounds gave a statistically significant reduction in the percentage of cells with a positive reaction to PCNA and Ki 67 antigen. TGF-β1, isotretinoin and tretinoin added to the culture resulted in the lowest percentage of PCNA positive cells. However, the lowest fraction of Ki 67 positive cells was observed after addition of isotretinoin. The obtained results also confirm the fact that the well-known regulatory proteins Bcl-2 and p53 play an important role in the regulation of apoptosis in the MCF-7 cell line, with lowered Bcl-2 expression accompanying easier apoptotic induction. The majority of the examined compounds act via the p53 pathway although some bypass this important proapoptotic factor.
Physical description
  • Department of Gynaecological Endocrinology, Medical University of Białystok, Białystok, Poland
  • Department of Gynaecological Endocrinology, Medical University of Białystok, Białystok, Poland
  • Department of Anatomy, Medical University of Białystok, Białystok, Poland
  • Department of Hematological Diagnostics, Medical University of Białystok, Białystok, Poland
  • Department of Gynaecological Endocrinology, Medical University of Białystok, Białystok, Poland
  • Arric BA, Korc M, Derynck R. (1990) Differential regulation of expression of three transforming growth factor beta species in human breast cancer cell lines by estradiol. Cancer Res.; 50: 299-303.
  • Attisan L, Wrana JL, Lopez-Caillas F, Massaque J. (1990) The transforming growth factor-β family. Annu Rev Cell Biol.; 6: 597-641.
  • Barnard JA, Lyons RM, Moses HL. (1990) The cell biology of transforming growth factor β. Biochem Biophys Acta.; 1032: 79-87.
  • Beenken SW, Hockett R Jr, Grizzle W, Weiss HE, Pickens A, Perloff M, Malone WF, Bland KI. (2002) Transforming growth factor-alpha: a surrogate endpoint biomarker? J Am Coll Surg.; 195: 149-58.
  • Birchenall-Roberts MC, Ruscetti FW, Kasper J, Lee HD, Friedman R, Geiser A, Sporn MB, Roberts AB, Kim SJ. (1995) Transcriptional regulation of the transforming growth factor β1 promoter by v-src gene products is mediated through the AP-1 complex. Mol Cell Biol.; 10: 4978-83.
  • Boise LH, Gonzalez-Garcia M, Postema ChE, Ding E, Lindsten T, Turka LA, Mao X, Nunez G, Thompson CB. (1993) bcl-x, a bcl-2-related gene that functions as a dominant regulator of apoptotic cell death. Cell.; 74: 597-608.
  • Bollag W, Peck R, Frey JR. (1992) Inhibition of proliferation by retinoids, cytokines and their combination in four human transformed epithelial cell lines. Cancer Lett.; 62: 167-72.
  • Chambon P. (1995) The molecular and genetic dissection of the retinoid signalling pathway. Rec Prog Horm Res.; 50: 317-32.
  • Chen H, Tritton TR, Kenny N, Absher M, Chiu JF. (1996) Tamoxifen induces TGF-beta 1 activity and apoptosis of human MCF-7 breast cancer cells in vitro. J Cell Biochem.; 61: 9-17.
  • Colletta AA, Wakefield LM, Howell FV, van Roozendaal KE, Danielpour D, Ebbs SR, Sporn MB, Baum M. (1990) Anti-oestrogens induce the secretion of active transforming growth factor β from human fetal fibroblasts. Br J Cancer.; 62: 405-9.
  • Coradini D, Biffi A, Pellizzaro C, Pironello E, Di Fronzo G. (1997) Combined effect of tamoxifen or interferon β and 4-hydroxyphenylretinamide on the growth of breast cancer cell lines. Tumour Biol.; 18: 22-9.
  • Cupp AS, Dufour JM, Kim G, Skinner MK, Kim KH. (1999) Action of retinoids on embryonic and early postnatal testis development. Endocrinology.; 140: 2343-52.
  • Czeczuga-Semeniuk E, Wołczyński S, Dzięcioł J, Dąbrowska M, Anchim T, Tomaszewska I. (2001) 13-cis retinoic acid and all-trans retinoic acid in regulation of the proliferation and survival of human breast cancer cells line MCF-7. Cell Mol Biol Lett.; 6: 925-39.
  • Danielpour D. (1996) Induction of transforming growth factor-beta autocrine activity by all trans-retinoic acid and 1 alpha 25-dihydroxyvitamin D3 in NRP-152 rat prostatic epithelial cells. J Cell Physiol.; 166: 231-9.
  • Desruisseau S, Ghazarossian-Ragni E, Chinot O, Martin PM. (1996) Divergent effect of TGF beta1 on growth and proteolytic modulation of human prostatic cancer cell lines. Int J Cancer.; 66: 796-801.
  • Durand B, Saunders M, Leroy P, Leid M, Chambon P. (1992) All-trans and 9-cis retinoic acid induction of CRABP II transcription is mediated by RAR-RXR heterodimers bound to DR1 and DR1 repeated motifs. Cell.; 71: 73-85.
  • Frey JR, Peck R, Bollag W. (1991) Antiproliferative activity of retinoids, interferon α and their combination in five human transformed cell lines. Cancer Lett.; 57: 223-7.
  • Glick AB, Flanders KC, Danielpour D, Yuspa SH, Sporn MB. (1989) Retinoic acid induces transforming growth factor-β2 in cultured keratinocytes and mouse epidermis. Cell Regul.; 1: 87-97.
  • Gross A, McDonnell JM, Korsmeyer SJ. (1999) BCL-2 family members and the mitochondria in apoptosis. Genes Dev.; 13: 1899-911.
  • Gucev ZS, Oh Y, Kelley KM, Rosenfeld RG. (1996) Insuline-like growth factor binding protein-3 mediates retinoic-acid and transforming growth factor β2-induced growth inhibition in human breast cancer cells. Cancer Res.; 56: 1545-50.
  • Hietanen S, Auvinen E, Syrjanen K, Syrjanen S. (1998) Anti-proliferative effect of retinoids and interferon-alpha-2a on vaginal cell lines derived from squamous intra-epithelial lesions. Int J Cancer.; 78: 338-45.
  • Hishikawa K, Oemar BS, Tanner FC, Nakaki T, Luscher TF, Fujii T. (1999) Connective tissue growth factor induces apoptosis in human breast cancer cell line MCF-7. J Biol Chem.; 274: 37461-6.
  • Jakowlew SB, Moody TW, Mariano JM. (1997) Transforming growth factor-beta receptors in human cancer cell lines: analysis of transcript, protein and proliferation. Anticancer Res.; 17: 1849-60.
  • Jeng MH, ten Dijke P, Iwata KK, Jordan VC. (1993) Regulation of the levels of three transforming growth factor β mRNAs by estrogen and their effects on the proliferation of human breast-cancer cells. Mol Cell Endocrinol.; 97: 115-23.
  • Kalkhoven E, Roelen BAJ, de Winter JP, Mummery CL, van den Eijnden-van Raaij AJM, van der Saag PT, van der Burg B. (1995) Resistance to transforming growth factor β and activin due to reduced receptor expression in human breast tumour cell lines. Cell Growth Diff.; 6: 1151-61.
  • Karey KP, Sirbascu DA. (1988) Differential responsiveness of human breast cancer cell lines MCF-7 and T47D to growth factors and 17β estradiol. Cancer Res.; 48: 4083-92.
  • Kim DH, Chang JH, Lee KH, Lee HY, Kim SJ. (1997) Mechanism of E1A-induced transforming growth factor-β (TGF-β) resistance in mouse keratinocytes involves repression of TGF-β type II transcription. J Biol Chem.; 272: 688-94.
  • Knabbe C, Lippman ME, Wakefield LM, Flanders KC, Kasiol A, Derynck R, Dickson RB. (1987) Evidence that transforming growth factor-β is a hormonally regulated negative growth factor β in human breast-cancer cells. Cell.; 48: 417-28.
  • Koli KM, Ramsey TT, Ko Y, Dugger TC, Brattain MG, Arteaga CL. (1997) Blockade of transforming growth factor-β signalling does not abrogate antiestrogen-induced growth inhibition of human breast carcinoma cells. J Biol Chem.; 272: 8296-302.
  • Lafon C, Mazars P, Guerrin M, Barboule N, Charcosset JY, Valette A. (1995) Early gene responses associated with transforming growth factor-beta 1 growth inhibition and autoinduction in MCF-7 breast adenocarcinoma cells. Biochim Biophys Acta.; 1266: 288-95.
  • Lauritsen KJ, List HJ, Reiter R, Wellstein A, Riegel AT. (2002) A role for TGF-β in estrogen and retinoid mediated regulation of the nuclear receptor coactivator AIB1 in MCF-7 breast cancer cells. Oncogene.; 21: 7147-55.
  • Liss C, Feket MJ, Hasina R, Lingen MW. (2002) Retinoic acid modulates the ability of macrophages to participate in the induction of the angiogenic phenotype in head and neck squamous cell carcinoma. Int J Cancer.; 100: 283-9.
  • Liu M, Iavarone A, Freedman LP. (1996) Transcriptional activation of the human p21 WAFI/CIFI gene by retinoic acid receptor. J Biol Chem.; 271: 31723-8.
  • Liu Y, Zhong X, Li W, Brattain MG, Banerji SS. (2000) The role of Sp1 in the differential expression of transforming growth factor beta receptor type II in human breast adenocarcinoma MCF-7 cells. J Biol Chem.; 275: 12231-6.
  • Mangiarotti R, Danova M, Alberici R, Pellicciari C. (1998) All-trans retinoic acid (ATRA)-induced apoptosis is preceded by G1 arrest in human MCF-7 breast cancer cells. Br J Cancer.; 77: 186-91.
  • Markowitz S, Wang J, Myeroff L, Parsons R, Sun L, Lutterbaugh J, Fan RS, Zborowska E, Kinzler KW, Vogelstein B, Brattain M, Willson JKV. (1995) Inactivation of the type II TGF-β receptor in colon cancer cells with microsatellite instability. Science.; 268: 1336-8.
  • Massaque J, Cheifetz S, Laiho M, Ralph DA. (1992) Transforming growth factor-beta. Cancer Surv.; 12: 81-103.
  • Mazars P, Barboule N, Baldin V, Vidal S, Ducommun B, Valette A. (1995) Effects of TGF-βeta 1 (transforming growth factor-beta 1) on the cell cycle regulation of human breast adenocarcinoma (MCF-7) cells. FEBS Lett.; 362: 295-300.
  • Nagy L, Thomazy VA, Shipley GL, Fesus L, Lamph W, Heyman RA, Chandraratna RAS, Davies PJA. (1995) Activation of retinoid X receptors induces apoptosis in HL-60 cell lines. Mol Cell Biol.; 15: 3540-51.
  • Niu MY, Ménard M, Reed JC, Krajewski S, Pratt MAC. (2001) Ectopic expression of cyclin D1 amplifies a retinoic acid-induced mitochondrial death pathway in breast cancer cells. Oncogene.; 20: 3506-18.
  • Perry RR, Kang Y, Greaves BR. (1995) Relationship between tamoxifen-induced transforming growth factor beta 1 expression, cytostasis and apoptosis in human breast cancer cells. Br J Cancer.; 72: 1441-6.
  • Pierce DF Jr, Gorska AE, Chytil A, Meise KS, Page DL, Coffey RJ Jr, Moses HL. (1995) Mammary tumor suppression by transforming growth factor beta 1 transgene expression. Proc Natl Acad Sci USA.; 92: 4254-8.
  • Reiss M. (1999) TGF-β and cancer. Microbes Infect.; 1: 1327-47.
  • Reiss M, Barcellos-Hoff MH. (1997) Transforming growth factor-β in breast cancer working hypothesis. Breast Cancer Res Treat.; 45: 81-95.
  • Roberts AB, Sporn MB. (1992) Mechanistic interrelationships between two superfamilies: the steroid/retinoid receptors and transforming growth factor β. Cancer Surv.; 14: 205-20.
  • Schadendorf D, Kern MA, Artuc M, Pahl HL, Rosenbach T, Fichtner I, Nurnberg W, Stuting S, Stebut E, Worm M, Makki A, Jurgovsky K, Kolde G, Henz BM. (1996) Treatment of melanoma cells with the synthetic retinoid CD-437 induces apoptosis via activation of AP-1 in vitro and causes growth inhibition in xenografts in vivo. J Cell Biol.; 135: 1889-98.
  • Shao ZM, Dawson MI, Li XS, Rishi AK, Sheikh MS, Han QX, Ordonez JV, Shroot B, Fontana JA. (1995) p53-independent G0/G1 arrest and apoptosis induced by a novel retinoid in human breast cancer cells. Oncogene.; 11: 493-504.
  • Sheikh MS, Shao ZM, Li XS, Ordonez JV, Conley BA, Wu S, Dowson MI, Han QX, Chao W, Quick T, Niles RN, Fontana JA. (1995) N-(4-hydroxyphenyl) retinamide (4HRP)-mediated biological actions involve retinoid receptor-independent pathways in human breast carcinoma. Carcinogenesis.; 16: 2477-86.
  • Smith WC, Nakshatri H, Leroy P, Rees J, Chambon P. (1991) A retinoic acid response element is present in the mouse cellular retinol binding protein I (mCRBP I) promoter. EMBO J.; 10: 2223-30.
  • Sporn MB, Roberts AB. (1991) Interactions of retinoids and transforming growth factor-β in regulation of cell differentiation and proliferation. Mol Endocrinol.; 5: 3-7.
  • Sporn MB, Roberts AB. (1992) Transforming growth factor-β: recent progress and new challenges. J Cell Biol.; 119: 1017-21.
  • Stewart AJ, Westley BR, May FEB. (1992) Modulation of the proliferative response of breast cancer cells to growth factors by oestrogen. Br J Cancer.; 66: 640-8.
  • Stoica A, Secada M, Fakhro A, Solomon HB, Fenster BD, Martin MB. (1997) The role of transforming growth factor-beta in the regulation of estrogen receptor expression in the MCF-7 breast cancer cell line. Endocrinology.; 138: 1498-505.
  • Sun L, Wu G, Willson JKV, Zborowska E, Yang J, Rajkarunanayake I, Wang J, Gentry LE, Wang XF, Brattain MG. (1994) Expression of transforming growth factor β type II receptor leads to reduced malignancy in human breast cancer MCF-7 cells. J Biol Chem.; 269: 26449-55.
  • Teixeira C, Pratt MAC. (1997) CDK2 is a target for retinoic acid-mediated growth inhibition in MCF-7 human breast cancer cells. Mol Endocrinol.; 11: 1191-202.
  • Toma S, Isnardi I, Raffo P, Dastoli G, De Francisci E, Riccardi L, Palumbo K, Bollag W. (1997) Effects of all-trans-retinoic acid and 13-cis-retinoic acid on breast cancer cell lines: growth inhibition and apoptosis induction. Int J Cancer.; 70: 619-27.
  • Zujewski JA, Vaughn-Cooke A, Flanders KC, Eckhaus MA, Lubet RA, Wakefield LM. (2001) Transforming growth factors-beta are not good biomarkers of chemopreventive efficacy in a preclinical breast cancer model system. Breast Cancer Res.; 3: 66-75.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.