Abstract
Purpose
Estrogen receptor α (ERα) mediates the growth stimulation of estrogen in breast cancer cells and is a useful predictive factor for response to endocrine therapy. It is reported that ERα was induced in ERα negative breast cancer cells by both DNA methyltransferase-1 (DNMT1) inhibitor 5-aza-2′-deoxycytidine (AZA) and histone deacetylase (HDAC) inhibitor trichostatin A (TSA). However, whether the breast cancer cells with induced ERα restore response to endocrine therapy requires to be further researched.
Patients and methods
Reverse transcriptase-polymerase chain reaction (RT-PCR) method was used to explore the change in the mRNA of ERα, PR and pS2 in the ERα negative breast cancer cells MDA-MB-435 treated with two chemicals (AZA + TSA). Water-soluble tetrazolium salt-8 (WST-8) method was used to study the proliferation rate of the breast cancer cells. Flow cytometer (FCW) was used to analyze the distribution of cell cycle of these breast cancer cells. Some xenograft models in nude mice were used to further study the results we found in vitro.
Results
In this study we observed that the mRNA of ERα, PR and pS2 in the ERα negative breast cancer cells MDA-MB-435 was re-expressed by treatment with AZA + TSA. The proliferation assay analysis showed AZA + TSA suppressed the proliferation of MDA-MB-435 cells, which were further suppressed by addition of 4-OH Tamoxifen (4-OHT). On the contrary, the proliferation of cells treated with 4-OHT alone showed no difference compared with the vehicle control. Cell cycle analysis showed AZA + TSA treated cells showed S phase arrest, which was partially attenuated by addition of estradiol (E2); furthermore, the effect of E2 on stimulation of cell cycle could be reversed by 4-OHT in the treated cells with induced ERα. In vivo experiment xenograft volume of MDA-MB-435 cells treated with AZA + TSA was smaller than that of the control (P < 0.01), and the xenograft of AZA + TSA treated cells was further suppressed by ovarectomy (P < 0.01).
Conclusions
Our data indicate that DNMT1 inhibitor AZA and HDAC inhibitor TSA play important roles in restoring sensitivity of the ERα negative breast cancer cells to endocrine therapy in vitro and in vivo.
Similar content being viewed by others
References
Antequera F, Bird A (1993) Number of CpG islands and genes in human and mouse. Proc Natl Acad Sci USA 90(24):11995–11999
Baylin S, Bestor TH (2002) Altered methylation patterns in cancer cell genomes: cause or consequence? Cancer Cell 1(4):299–305
Bovenzi V, Momparler RL (2001) Antineoplastic action of 5-aza-2′-deoxycytidine and histone deacetylase inhibitor and their effect on the expression of retinoic acid receptor beta and estrogen receptor alpha genes in breast carcinoma cells. Cancer Chemother Pharmacol 48(1):71–76
Butler LM, Agus DB, Scher HI, Higgins B, Rose A, Cordon-Cardo C, Thaler HT, Rifkind RA, Marks PA, Richon VM (2000) Suberoylanilide hydroxamic acid, an inhibitor of histone deacetylase, suppresses the growth of prostate cancer cells in vitro and in vivo. Cancer Res 60(18):5165–5170
Cameron EE, Bachman KE, Myohanen S, Herman JG, Baylin SB (1999) Synergy of demethylation and histone deacetylase inhibition in the re-expression of genes silenced in cancer. Nat Genet 21(1):103–107
Cheung WL, Briggs SD, Allis CD (2000) Acetylation and chromosomal functions. Curr Opin Cell Biol 12(3):326–333
Clark CM (2000) Prognostic and predictive factors. In: Harris JR, Lippman ME, Morrow M, Osborne CK (eds) Diseases of the breast. 2 edn. Lippincott Williams & Wilkins, Philadelphia, pp 489–524
Ferguson AT, Lapidus RG, Baylin SB, Davidson NE (1995) Demethylation of the estrogen receptor gene in estrogen receptor-negative breast cancer cells can reactivate estrogen receptor gene expression. Cancer Res 55(11):2279–2283
Giacinti L, Claudio PP, Lopez M, Giordano A (2006) Epigenetic information and estrogen receptor alpha expression in breast cancer. Oncologist 11(1):1–8
Hedden A, Muller V, Jensen EV (1995) A new interpretation of antiestrogen action. Ann N Y Acad Sci 761:109–120
Jang ER, Lim SJ, Lee ES, Jeong G, Kim TY, Bang YJ, Lee JS (2004) The histone deacetylase inhibitor trichostatin A sensitizes estrogen receptor alpha-negative breast cancer cells to tamoxifen. Oncogene 23(9):1724–1736
Jones PA, Baylin SB (2002) The fundamental role of epigenetic events in cancer. Nat Rev Genet 3(6):415–428
Jordan VC (2001) The past, present, and future of selective estrogen receptor modulation. Ann N Y Acad Sci 949:72–79
Keen JC, Yan L, Mack KM, Pettit C, Smith D, Sharma D, Davidson NE (2003a) A novel histone deacetylase inhibitor, scriptaid, enhances expression of functional estrogen receptor alpha (ER) in ER negative human breast cancer cells in combination with 5-aza 2′-deoxycytidine. Breast Cancer Res Treat 81(3):177–186
Keen JC, Yan L, Mack KM, Pettit C, Smith D, Sharma D, Davidson NE (2003b) A novel histone deacetylase inhibitor, scriptaid, enhances expression of functional estrogen receptor alpha (ER) in ER negative human breast cancer cells in combination with 5-aza 2′-deoxycytidine. Breast Cancer Res Treat 81(3):177–186
Kumar R, Wang RA, Mazumdar A, Talukder AH, Mandal M, Yang Z, Bagheri-Yarmand R, Sahin A, Hortobagyi G, Adam L et al (2002) A naturally occurring MTA1 variant sequesters oestrogen receptor-alpha in the cytoplasm. Nature 418(6898):654–657
Langsenlehner U, Krippl P, Renner W, Yazdani-Biuki B, Wolf G, Wascher TC, Paulweber B, Weitzer W, Samonigg H (2003) The common 677C > T gene polymorphism of methylenetetrahydrofolate reductase gene is not associated with breast cancer risk. Breast Cancer Res Treat 81(2):169–172
Marks PA, Richon VM, Rifkind RA (2000) Histone deacetylase inhibitors: inducers of differentiation or apoptosis of transformed cells. J Natl Cancer Inst 92(15):1210–1216
Munster PN, Troso-Sandoval T, Rosen N, Rifkind R, Marks PA, Richon VM (2001) The histone deacetylase inhibitor suberoylanilide hydroxamic acid induces differentiation of human breast cancer cells. Cancer Res 61(23):8492–8497
Segura-Pacheco B, Trejo-Becerril C, Perez-Cardenas E, Taja-Chayeb L, Mariscal I, Chavez A, Acuna C, Salazar AM, Lizano M, Duenas-Gonzalez A (2003) Reactivation of tumor suppressor genes by the cardiovascular drugs hydralazine and procainamide and their potential use in cancer therapy. Clin Cancer Res 9(5):1596–1603
Shou J, Massarweh S, Osborne CK, Wakeling AE, Ali S, Weiss H, Schiff R (2004) Mechanisms of tamoxifen resistance: increased estrogen receptor-HER2/neu cross-talk in ER/HER2-positive breast cancer. J Natl Cancer Inst 96(12):926–935
Toth KF, Knoch TA, Wachsmuth M, Frank-Stohr M, Stohr M, Bacher CP, Muller G, Rippe K (2004) Trichostatin A-induced histone acetylation causes decondensation of interphase chromatin. J Cell Sci 117(Pt 18):4277–4287
Touma SE, Goldberg JS, Moench P, Guo X, Tickoo SK, Gudas LJ, Nanus DM (2005) Retinoic acid and the histone deacetylase inhibitor trichostatin a inhibit the proliferation of human renal cell carcinoma in a xenograft tumor model. Clin Cancer Res 11(9):3558–3566
Vigushin DM, Ali S, Pace PE, Mirsaidi N, Ito K, Adcock I, Coombes RC (2001) Trichostatin A is a histone deacetylase inhibitor with potent antitumor activity against breast cancer in vivo. Clin Cancer Res 7(4):971–976
Yang X, Ferguson AT, Nass SJ, Phillips DL, Butash KA, Wang SM, Herman JG, Davidson NE (2000) Transcriptional activation of estrogen receptor alpha in human breast cancer cells by histone deacetylase inhibition. Cancer Res 60(24):6890–6894
Yang X, Phillips DL, Ferguson AT, Nelson WG, Herman JG, Davidson NE (2001a) Synergistic activation of functional estrogen receptor (ER)-alpha by DNA methyltransferase and histone deacetylase inhibition in human ER-alpha-negative breast cancer cells. Cancer Res 61(19):7025–7029
Yang X, Phillips DL, Ferguson AT, Nelson WG, Herman JG, Davidson NE (2001b) Synergistic activation of functional estrogen receptor (ER)-alpha by DNA methyltransferase and histone deacetylase inhibition in human ER-alpha-negative breast cancer cells. Cancer Res 61(19):7025–7029
Zhu K, Davidson NE, Hunter S, Yang X, Payne-Wilks K, Roland CL, Phillips D, Bentley C, Dai M, Williams SM (2003) Methyl-group dietary intake and risk of breast cancer among African–American women: a case-control study by methylation status of the estrogen receptor alpha genes. Cancer Causes Control 14(9):827–836
Acknowledgments
This research was supported in part by the Outstanding Young Investigator Award of National Natural Science Foundation of China (30025015), National Key Project of China (2001BA703BO5), National Natural Science Foundation of China (30371580, 30572109, 30570695) and the Grant from Shanghai Science and Technology Committee (03J14019).
Author information
Authors and Affiliations
Corresponding author
Additional information
J. Fan, W.-J. Yin and J.-S. Lu contributed equally to the work.
Rights and permissions
About this article
Cite this article
Fan, J., Yin, WJ., Lu, JS. et al. ERα negative breast cancer cells restore response to endocrine therapy by combination treatment with both HDAC inhibitor and DNMT inhibitor. J Cancer Res Clin Oncol 134, 883–890 (2008). https://doi.org/10.1007/s00432-008-0354-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00432-008-0354-x