Abstract
Majority of women with estrogen receptor (ER)-positive breast cancers initially respond to hormone therapies such as tamoxifen (TAM; antagonist of estrogen). However, many tumors eventually become resistant to TAM. Therefore, understanding the various cellular components involved in causing resistance to TAM is of paramount importance in designing novel entities for efficacious hormone therapy. Previously, we found that suppression of BIK gene expression induced TAM resistance in MCF-7 breast cancer cells. In order to understand the response of these cells to TAM and its association with resistance, a microarray analysis of gene expression was performed in the BIK-suppressed MCF-7 cells and compared it to the TAM-only-treated cells (controls). Several genes participating in various cellular pathways were identified. Molecules identified in the drug resistance pathway were 14-3-3z or YWHAZ, WEE1, PRKACA, NADK, and HSP90AA 1. Further, genes involved in cell cycle control, apoptosis, and cell proliferation were also found differentially expressed in these cells. Transcriptional and translational analysis of key molecules such as STAT2, AKT 3, and 14-3-3z revealed similar changes at the messenger RNA (mRNA) as well as at the protein level. Importantly, there was no cytotoxic effect of TAM on BIK-suppressed MCF-7 cells. Further, these cells were not arrested at the G0-G1 phase of the cell cycle although 30 % of BIK-suppressed cells were arrested at the G2 phase of the cycle on TAM treatment. Furthermore, we found a relevant interaction between 14-3-3z and WEE1, suggesting that the cytotoxic effect of TAM was prevented in BIK-suppressed cells because this interaction leads to transitory arrest in the G2 phase leading to the repair of damaged DNA and allowing the cells to proliferate.
Similar content being viewed by others
References
Gherardi E, Rota E, Calandra S, Genova R, Tamborino A. Relationship among the concentrations of serum lipoproteins and changes in their chemical composition in patients with untreated nephrotic syndrome. Eur J Clin Invest. 1977;7(6):563–70.
Vargo-Gogola T, Rosen JM. Modelling breast cancer: one size does not fit all. Nat Rev Cancer. 2007;7(9):659–72. doi:10.1038/nrc2193.
Jordan VC, Murphy CS. Endocrine pharmacology of antiestrogens as antitumor agents. Endocr Rev. 1990;11(4):578–610.
Sheen YY, Ruh TS, Mangel WF, Katzenellenbogen BS. Antiestrogenic potency and binding characteristics of the triphenylethylene H1285 in MCF-7 human breast cancer cells. Cancer Res. 1985;45(9):4192–9.
Gee JM, Harper ME, Hutcheson IR, Madden TA, Barrow D, Knowlden JM, et al. The antiepidermal growth factor receptor agent gefitinib (ZD1839/Iressa) improves antihormone response and prevents development of resistance in breast cancer in vitro. Endocrinology. 2003;144(11):5105–17. doi:10.1210/en.2003-0705en.2003-0705.
Hoskins JM, Carey LA, McLeod HL. CYP2D6 and tamoxifen: DNA matters in breast cancer. Nat Rev Cancer. 2009;9(8):576–86. doi:10.1038/nrc2683nrc2683.
Gutierrez MC, Detre S, Johnston S, Mohsin SK, Shou J, Allred DC, et al. Molecular changes in tamoxifen-resistant breast cancer: relationship between estrogen receptor, HER-2, and p38 mitogen-activated protein kinase. J Clin Oncol. 2005;23(11):2469–76. doi:10.1200/JCO.2005.01.172.
Shi L, Dong B, Li Z, Lu Y, Ouyang T, Li J, et al. Expression of ER-{alpha}36, a novel variant of estrogen receptor {alpha}, and resistance to tamoxifen treatment in breast cancer. J Clin Oncol. 2009;27(21):3423–9. doi:10.1200/JCO.2008.17.2254JCO.2008.17.2254.
Honma N, Horii R, Iwase T, Saji S, Younes M, Takubo K, et al. Clinical importance of estrogen receptor-beta evaluation in breast cancer patients treated with adjuvant tamoxifen therapy. J Clin Oncol. 2008;26(22):3727–34. doi:10.1200/JCO.2007.14.296826/22/3727.
Viedma-Rodriguez R, Baiza-Gutman L, Salamanca-Gomez F, Diaz-Zaragoza M, Martinez-Hernandez G, Ruiz Esparza-Garrido R, et al. Mechanisms associated with resistance to tamoxifen in estrogen receptor-positive breast cancer (review). Oncol Rep. 2014. doi:10.3892/or.2014.3190.
Viedma-Rodriguez R, Baiza-Gutman LA, Garcia-Carranca A, Moreno-Fierros L, Salamanca-Gomez F, Arenas-Aranda D. Suppression of the death gene BIK is a critical factor for resistance to tamoxifen in MCF-7 breast cancer cells. Int J Oncol. 2013;43(6):177786. doi:10.3892/ijo.2013.2127.
Lopez J, Hesling C, Prudent J, Popgeorgiev N, Gadet R, Mikaelian I, et al. Src tyrosine kinase inhibits apoptosis through the Erk1/2-dependent degradation of the death accelerator Bik. Cell Death Differ. 2012;19(9):1459–69. doi:10.1038/cdd.2012.21cdd201221.
Fu Y, Li J, Lee AS. GRP78/BiP inhibits endoplasmic reticulum BIK and protects human breast cancer cells against estrogen starvation-induced apoptosis. Cancer Res. 2007;67(8):3734–40. doi:10.1158/0008-5472.CAN-06-4594.
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680–5.
Schaefer MH, Fontaine JF, Vinayagam A, Porras P, Wanker EE, Andrade-Navarro MA. HIPPIE: integrating protein interaction networks with experiment based quality scores. PLoS One. 2012;7(2):e31826. doi:10.1371/journal.pone.0031826PONE-D-11-20814.
Shannon P, Markiel A, Ozier O, Baliga NS, Wang JT, Ramage D, et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res. 2003;13(11):2498–504. doi:10.1101/gr.123930313/11/2498.
Rajput S, Kumar BN, Sarkar S, Das S, Azab B, Santhekadur PK, et al. Targeted apoptotic effects of thymoquinone and tamoxifen on XIAP mediated Akt regulation in breast cancer. PLoS One. 2013;8(4):e61342. doi:10.1371/journal.pone.0061342PONE-D-12-38938.
Pouliot LM, Chen YC, Bai J, Guha R, Martin SE, Gottesman MM, et al. Cisplatin sensitivity mediated by WEE1 and CHK1 is mediated by miR-155 and the miR-15 family. Cancer Res. 2012;72(22):5945–55. doi:10.1158/0008-5472.CAN-12-14000008-5472.CAN-12-1400.
Chu SH, Liu YW, Zhang L, Liu B, Li L, Shi JZ. Regulation of survival and chemoresistance by HSP90AA1 in ovarian cancer SKOV3 cells. Mol Biol Rep. 2013;40(1):1–6. doi:10.1007/s11033-012-1930-3.
Moody SE, Schinzel AC, Singh S, Izzo F, Strickland MR, Luo L, et al. PRKACA mediates resistance to HER2-targeted therapy in breast cancer cells and restores anti-apoptotic signaling. Oncogene. 2014. doi:10.1038/onc.2014.153onc2014153.
Carraway 3rd KL. E3 ubiquitin ligases in ErbB receptor quantity control. Semin Cell Dev Biol. 2010;21(9):936–43. doi:10.1016/j.semcdb.2010.09.006S1084-9521(10)00153-9.
Liu B, Wen X, Cheng Y. Survival or death: disequilibrating the oncogenic and tumor suppressive autophagy in cancer. Cell Death Dis. 2013;4:e892. doi:10.1038/cddis.2013.422cddis2013422.
Privette LM, Petty EM. CHFR: a novel mitotic checkpoint protein and regulator of tumorigenesis. Transl Oncol. 2008;1(2):57–64.
Jordan NJ, Dutkowski CM, Barrow D, Mottram HJ, Hutcheson IR, Nicholson RI, et al. Impact of dual mTORC1/2 mTOR kinase inhibitor AZD8055 on acquired endocrine resistance in breast cancer in vitro. Breast Cancer Res. 2014;16(1):R12. doi:10.1186/bcr3604bcr3604.
Bergamaschi A, Christensen BL, Katzenellenbogen BS. Reversal of endocrine resistance in breast cancer: interrelationships among 14-3-3zeta, FOXM1, and a gene signature associated with mitosis. Breast Cancer Res. 2011;13(3):R70. doi:10.1186/bcr2913bcr2913.
Frasor J, Chang EC, Komm B, Lin CY, Vega VB, Liu ET, et al. Gene expression preferentially regulated by tamoxifen in breast cancer cells and correlations with clinical outcome. Cancer Res. 2006;66(14):7334–40. doi:10.1158/0008-5472.CAN-05-4269.
Zeitz MJ, Ay F, Heidmann JD, Lerner PL, Noble WS, Steelman BN, et al. Genomic interaction profiles in breast cancer reveal altered chromatin architecture. PLoS One. 2013;8(9):e73974. doi:10.1371/journal.pone.0073974PONE-D-13-13446.
Sun M, Gomes S, Chen P, Frankenberger CA, Sankarasharma D, Chung CH, et al. RKIP and HMGA2 regulate breast tumor survival and metastasis through lysyl oxidase and syndecan-2. Oncogene. 2014;33(27):3528–37. doi:10.1038/onc.2013.328onc2013328.
Dutta S, Bandyopadhyay C, Bottero V, Veettil MV, Wilson L, Pins MR, et al. Angiogenin interacts with the plasminogen activation system at the cell surface of breast cancer cells to regulate plasmin formation and cell migration. Mol Oncol. 2014;8(3):483–507. doi:10.1016/j.molonc.2013.12.017S1574-7891(13)00189-0.
Nagashima K, Shumway SD, Sathyanarayanan S, Chen AH, Dolinski B, Xu Y, et al. Genetic and pharmacological inhibition of PDK1 in cancer cells: characterization of a selective allosteric kinase inhibitor. J Biol Chem. 2011;286(8):6433–48. doi:10.1074/jbc.M110.156463M110.156463.
Gamero AM, Young MR, Mentor-Marcel R, Bobe G, Scarzello AJ, Wise J, et al. STAT2 contributes to promotion of colorectal and skin carcinogenesis. Cancer Prev Res (Phila). 2010;3(4):495–504. doi:10.1158/1940-6207.CAPR-09-01051940-6207.CAPR-09-0105.
Chin YR, Yoshida T, Marusyk A, Beck AH, Polyak K, Toker A. Targeting Akt3 signaling in triple-negative breast cancer. Cancer Res. 2014;74(3):964–73. doi:10.1158/0008-5472.CAN-13-21750008-5472.CAN-13-2175.
Rehman SK, Li SH, Wyszomierski SL, Wang Q, Li P, Sahin O, et al. 14-3-3zeta orchestrates mammary tumor onset and progression via miR-221-mediated cell proliferation. Cancer Res. 2014;74(1):363–73. doi:10.1158/0008-5472.CAN-13-20160008-5472.CAN-13-2016.
Neal CL, Yao J, Yang W, Zhou X, Nguyen NT, Lu J, et al. 14-3-3zeta overexpression defines high risk for breast cancer recurrence and promotes cancer cell survival. Cancer Res. 2009;69(8):3425–32. doi:10.1158/0008-5472.CAN-08-27650008-5472.CAN-08-2765.
Ballesta A, Lopez J, Popgeorgiev N, Gonzalo P, Doumic M, Gillet G. Data-driven modeling of SRC control on the mitochondrial pathway of apoptosis: implication for anticancer therapy optimization. PLoS Comput Biol. 2013;9(4):e1003011. doi:10.1371/journal.pcbi.1003011PCOMPBIOL-D-12-01842.
McGlynn LM, Kirkegaard T, Edwards J, Tovey S, Cameron D, Twelves C, et al. Ras/Raf-1/MAPK pathway mediates response to tamoxifen but not chemotherapy in breast cancer patients. Clin Cancer Res. 2009;15(4):1487–95. doi:10.1158/1078-0432.CCR-07-496715/4/1487.
Sanchez-Munoz A, Gallego E, de Luque V, Perez-Rivas LG, Vicioso L, Ribelles N, et al. Lack of evidence for KRAS oncogenic mutations in triple-negative breast cancer. BMC Cancer. 2010;10:136. doi:10.1186/1471-2407-10-1361471-2407-10-136.
Roy S, McPherson RA, Apolloni A, Yan J, Lane A, Clyde-Smith J, et al. 14-3-3 facilitates Ras-dependent Raf-1 activation in vitro and in vivo. Mol Cell Biol. 1998;18(7):3947–55.
Leung EY, Kim JE, Askarian-Amiri M, Joseph WR, McKeage MJ, Baguley BC. Hormone resistance in Two MCF-7 breast cancer cell lines is associated with reduced mTOR signaling, decreased glycolysis, and increased sensitivity to cytotoxic drugs. Front Oncol. 2014;4:221. doi:10.3389/fonc.2014.00221.
Faridi J, Wang L, Endemann G, Roth RA. Expression of constitutively active AKT3 in MCF-7 breast cancer cells reverses the estrogen and tamoxifen responsivity of these cells in vivo. Clin Cancer Res. 2003;9(8):2933–9.
Corum DG, Tsichlis PN, Muise-Helmericks RC. AKT3 controls mitochondrial biogenesis and autophagy via regulation of the major nuclear export protein CRM-1. FASEB J. 2014;28(1):395–407. doi:10.1096/fj.13-235382fj.13-235382.
Schoenlein PV, Periyasamy-Thandavan S, Samaddar JS, Jackson WH, Barrett JT. Autophagy facilitates the progression of ERalpha-positive breast cancer cells to antiestrogen resistance. Autophagy. 2009;5(3):400–3.
Hamilton DH, Huang B, Fernando RI, Tsang KY, Palena C. WEE1 inhibition alleviates resistance to immune attack of tumor cells undergoing epithelial-mesenchymal transition. Cancer Res. 2014;74(9):2510–9. doi:10.1158/0008-5472.CAN-13-18940008-5472.CAN-13-1894.
Danes CG, Wyszomierski SL, Lu J, Neal CL, Yang W, Yu D. 14-3-3 zeta down-regulates p53 in mammary epithelial cells and confers luminal filling. Cancer Res. 2008;68(6):1760–7. doi:10.1158/0008-5472.CAN-07-317768/6/1760.
Matta A, Siu KW, Ralhan R. 14-3-3 zeta as novel molecular target for cancer therapy. Expert Opin Ther Targets. 2012;16(5):515–23. doi:10.1517/14728222.2012.668185.
van Hemert MJ, Steensma HY, van Heusden GP. 14-3-3 proteins: key regulators of cell division, signalling and apoptosis. Bioessays. 2001;23(10):936–46. doi:10.1002/bies.1134.
Tzivion G, Gupta VS, Kaplun L, Balan V. 14-3-3 proteins as potential oncogenes. Semin Cancer Biol. 2006;16(3):203–13. doi:10.1016/j.semcancer.2006.03.004.
Coleman TR, Dunphy WG. Cdc2 regulatory factors. Curr Opin Cell Biol. 1994;6(6):877–82.
Lal S, Burkhart RA, Beeharry N, Bhattacharjee V, Londin ER, Cozzitorto JA, et al. HuR posttranscriptionally regulates WEE1: implications for the DNA damage response in pancreatic cancer cells. Cancer Res. 2014;74(4):1128–40. doi:10.1158/0008-5472.CAN-13-191574/4/1128.
Ghiasi N, Habibagahi M, Rosli R, Ghaderi A, Yusoff K, Hosseini A, et al. Tumour suppressive effects of WEE1 gene silencing in breast cancer cells. Asian Pac J Cancer Prev. 2013;14(11):6605–11.
Magnussen GI, Holm R, Emilsen E, Rosnes AK, Slipicevic A, Florenes VA. High expression of Wee1 is associated with poor disease-free survival in malignant melanoma: potential for targeted therapy. PLoS One. 2012;7(6):e38254. doi:10.1371/journal.pone.0038254PONE-D-12-03007.
Mir SE, De Witt Hamer PC, Krawczyk PM, Balaj L, Claes A, Niers JM, et al. In silico analysis of kinase expression identifies WEE1 as a gatekeeper against mitotic catastrophe in glioblastoma. Cancer Cell. 2010;18(3):244–57. doi:10.1016/j.ccr.2010.08.011S1535-6108(10)00308-9.
Acknowledgments
We thank Jorge Ramírez-Salcedo and collaborators for their technical assistance in the microarray analysis. This study was performed in partial fulfillment of the requirements for the Ph.D. degree in Biomedical Sciences of RV-R, who was the recipient of doctoral fellowship provided by CONACyT-México (grant no. 207148), at the Universidad Nacional Autónoma de México. This study was supported by grants Salud-2007-785-063 from CONACyT-México and PAPIIT, DGAPA, UNAM, project IN223014.
Conflicts of interest
None
Author information
Authors and Affiliations
Corresponding author
Additional information
Diego Arenas-Aranda passed away last 2014 (1958–2014).
Rights and permissions
About this article
Cite this article
Viedma-Rodríguez, R., Ruiz Esparza-Garrido, R., Baiza-Gutman, L.A. et al. Involvement of multiple cellular pathways in regulating resistance to tamoxifen in BIK-suppressed MCF-7 cells. Tumor Biol. 36, 6991–7005 (2015). https://doi.org/10.1007/s13277-015-3374-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13277-015-3374-6