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E2/ERβ Inhibits PPARα to Regulate Cell-Proliferation and Enhance Apoptosis in Hep3B-Hepatocellular Carcinoma

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Pathology & Oncology Research

Abstract

Peroxisome proliferator-activated receptor-α (PPARα) is a member of the nuclear receptor superfamily involved in hepatocarcinogenesis in rodents. In previous studies on liver tumor tissues, PPARα mRNA expression was found to be significantly higher and overexpression of ERα inhibited the PPARα expression, cell-proliferation and also induced apoptosis in Hep3B cell. However, the role of ERβ is not known yet. Therefore, the aim of this study is to define the role of ERβ on PPARα in Hep3B cells. The effect of PPARα signaling cascade were monitored by inducing Hep3B cells by fenofibrate. Further the cells were transfected with pCMV-ERβ and the consequences of ERβ-overexpression on the PPARα induced changes such as enhanced cell-proliferation and suppressed apoptosis were determined using western blot analysis and TUNEL assay. The EMSA was used to identify whether ERβ modulates PPARα expression by binding to PPARα promoter region to repress PPARα promoter activity. In addition, the direct interaction between ERβ and PPARα proteins was verified by co-immunoprecipitation assay. Our results show that the overexpressed ERβ not only attenuated the effects of fenofibrate to induce the levels of apoptosis protein such as Cyt.c, Caspase 9 and Caspase 3 but also inhibited the levels of survival protein such Bcl-xL, p-Bad, cyclin A and cyclin E. All these effects of E2/ERβ resulted in the enhancement of mitochondria dependent apoptotic pathway and the attenuation of cell proliferation. Moreover, the overexpressed ERβ reduced the mRNA and protein levels of PPARα and its downstream Acyl-CoA oxidase (ACO). EMSA results show that ERβ directly binds to PPRE and inhibit PPARα gene expression and according to immunoprecipitation assay ERβ also binds strongly with PPARα. The E2/ERβ further inhibited the fenofibrate-induced nuclear translocation of PPARα. Taken together, ERβ might directly downregulate PPARα gene expression and inhibit the nuclear translocation to suppress the proliferation and induce the apoptosis of Hep3B cells.

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Reference

  1. Bruix J, Boix L, Sala M, Llovet JM (2004) Focus on hepatocellular carcinoma. Cancer Cell 5(3):215–219

    Article  CAS  PubMed  Google Scholar 

  2. Reddy JK, Krishnakantha TP (1975) Hepatic peroxisome proliferation: induction by two novel compounds structurally unrelated to clofibrate. Science 190(4216):787–789

    Article  CAS  PubMed  Google Scholar 

  3. Yeldandi AV, Rao MS, Reddy JK (2000) Hydrogen peroxide generation in peroxisome proliferator-induced oncogenesis. Mutat Res 448(2):159–177

    Article  CAS  PubMed  Google Scholar 

  4. Wu CY, Huang HM, Cho DY (2015) An acute bleeding metastatic spinal tumor from HCC causes an acute onset of cauda equina syndrome. Biomedicine 5(3):18. doi:10.7603/s40681-015-0018-5

    Article  PubMed  PubMed Central  Google Scholar 

  5. Misra P, Reddy JK (2014) Peroxisome proliferator-activated receptor-alpha activation and excess energy burning in hepatocarcinogenesis. Biochimie 98:63–74. doi:10.1016/j.biochi.2013.11.011

    Article  CAS  PubMed  Google Scholar 

  6. Reddy JK, Azarnoff DL, Hignite CE (1980) Hypolipidaemic hepatic peroxisome proliferators form a novel class of chemical carcinogens. Nature 283(5745):397–398

    Article  CAS  PubMed  Google Scholar 

  7. Akriviadis EA, Llovet JM, Efremidis SC, Shouval D, Canelo R, Ringe B, Meyers WC (1998) Hepatocellular carcinoma. Br J Surg 85(10):1319–1331. doi:10.1046/j.1365-2168.1998.00865.x

    Article  CAS  PubMed  Google Scholar 

  8. Lin CY, Strom A, Li Kong S, Kietz S, Thomsen JS, Tee JB, Vega VB, Miller LD, Smeds J, Bergh J, Gustafsson JA, Liu ET (2007) Inhibitory effects of estrogen receptor beta on specific hormone-responsive gene expression and association with disease outcome in primary breast cancer. Breast Cancer Res BCR 9(2):R25. doi:10.1186/bcr1667

    Article  PubMed  Google Scholar 

  9. Lazennec G (2006) Estrogen receptor beta, a possible tumor suppressor involved in ovarian carcinogenesis. Cancer Lett 231(2):151–157. doi:10.1016/j.canlet.2005.01.021

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Treeck O, Pfeiler G, Horn F, Federhofer B, Houlihan H, Vollmer A, Ortmann O (2007) Novel estrogen receptor beta transcript variants identified in human breast cancer cells affect cell growth and apoptosis of COS-1 cells. Mol Cell Endocrinol 264(1–2):50–60. doi:10.1016/j.mce.2006.10.003

    Article  CAS  PubMed  Google Scholar 

  11. Behrens D, Gill JH, Fichtner I (2007) Loss of tumourigenicity of stably ERbeta-transfected MCF-7 breast cancer cells. Mol Cell Endocrinol 274(1–2):19–29. doi:10.1016/j.mce.2007.05.012

    Article  CAS  PubMed  Google Scholar 

  12. Paruthiyil S, Parmar H, Kerekatte V, Cunha GR, Firestone GL, Leitman DC (2004) Estrogen receptor beta inhibits human breast cancer cell proliferation and tumor formation by causing a G2 cell cycle arrest. Cancer Res 64(1):423–428

    Article  CAS  PubMed  Google Scholar 

  13. Strom A, Hartman J, Foster JS, Kietz S, Wimalasena J, Gustafsson JA (2004) Estrogen receptor beta inhibits 17beta-estradiol-stimulated proliferation of the breast cancer cell line T47D. Proc Natl Acad Sci U S A 101(6):1566–1571. doi:10.1073/pnas.0308319100

    Article  PubMed  PubMed Central  Google Scholar 

  14. Carruba G (2007) Estrogen and prostate cancer: an eclipsed truth in an androgen-dominated scenario. J Cell Biochem 102(4):899–911. doi:10.1002/jcb.21529

    Article  CAS  PubMed  Google Scholar 

  15. Stettner M, Kaulfuss S, Burfeind P, Schweyer S, Strauss A, Ringert RH, Thelen P (2007) The relevance of estrogen receptor-beta expression to the antiproliferative effects observed with histone deacetylase inhibitors and phytoestrogens in prostate cancer treatment. Mol Cancer Ther 6(10):2626–2633. doi:10.1158/1535-7163.MCT-07-0197

    Article  CAS  PubMed  Google Scholar 

  16. Cheng J, Lee EJ, Madison LD, Lazennec G (2004) Expression of estrogen receptor beta in prostate carcinoma cells inhibits invasion and proliferation and triggers apoptosis. FEBS Lett 566(1–3):169–172. doi:10.1016/j.febslet.2004.04.025

    Article  PubMed  Google Scholar 

  17. Chen GG, Zeng Q, Tse GM (2008) Estrogen and its receptors in cancer. Med Res Rev 28(6):954–974. doi:10.1002/med.20131

    Article  CAS  PubMed  Google Scholar 

  18. Jeong S, Yoon M (2007) Inhibition of the actions of peroxisome proliferator-activated receptor alpha on obesity by estrogen. Obesity 15(6):1430–1440. doi:10.1038/oby.2007.171

    Article  CAS  PubMed  Google Scholar 

  19. Huang EJ, Wu CC, Huang HP, Liu JY, Lin CS, Chang YZ, Lin JA, Lin JG, Chen LM, Lee SD, Kuo WW, Huang CY (2006) Apoptotic and anti-proliferative effects of 17beta-estradiol and 17beta-estradiol-like compounds in the Hep3B cell line. Mol Cell Biochem 290(1–2):1–7. doi:10.1007/s11010-005-9000-y

    Article  CAS  PubMed  Google Scholar 

  20. Marx N, Duez H, Fruchart JC, Staels B (2004) Peroxisome proliferator-activated receptors and atherogenesis: regulators of gene expression in vascular cells. Circ Res 94(9):1168–1178. doi:10.1161/01.RES.0000127122.22685.0A

    Article  CAS  PubMed  Google Scholar 

  21. Lee CH, Olson P, Evans RM (2003) Minireview: lipid metabolism, metabolic diseases, and peroxisome proliferator-activated receptors. Endocrinology 144(6):2201–2207. doi:10.1210/en.2003-0288

    Article  CAS  PubMed  Google Scholar 

  22. Lefebvre P, Chinetti G, Fruchart JC, Staels B (2006) Sorting out the roles of PPAR alpha in energy metabolism and vascular homeostasis. J Clin Invest 116(3):571–580. doi:10.1172/JCI27989

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Venalainen T, Molnar F, Oostenbrink C, Carlberg C, Perakyla M (2010) Molecular mechanism of allosteric communication in the human PPARalpha-RXRalpha heterodimer. Proteins 78(4):873–887. doi:10.1002/prot.22613

    Article  PubMed  Google Scholar 

  24. Ringseis R, Gutgesell A, Dathe C, Brandsch C, Eder K (2007) Feeding oxidized fat during pregnancy up-regulates expression of PPARalpha-responsive genes in the liver of rat fetuses. Lipids Health Dis 6:6. doi:10.1186/1476-511X-6-6

    Article  PubMed  PubMed Central  Google Scholar 

  25. Peters JM, Cheung C, Gonzalez FJ (2005) Peroxisome proliferator-activated receptor-alpha and liver cancer: where do we stand? J Mol Med 83(10):774–785. doi:10.1007/s00109-005-0678-9

    Article  CAS  PubMed  Google Scholar 

  26. Nishimura J, Dewa Y, Okamura T, Jin M, Saegusa Y, Kawai M, Umemura T, Shibutani M, Mitsumori K (2008) Role of Nrf2 and oxidative stress on fenofibrate-induced hepatocarcinogenesis in rats. Toxicological Sciences: an Official Journal of the Society of Toxicology 106(2):339–349. doi:10.1093/toxsci/kfn174

    Article  CAS  Google Scholar 

  27. Bisteau X, Caldez MJ, Kaldis P (2014) The complex relationship between liver cancer and the cell cycle: a story of multiple regulations. Cancer 6(1):79–111. doi:10.3390/cancers6010079

    Article  Google Scholar 

  28. Slingerland J, Pagano M (2000) Regulation of the cdk inhibitor p27 and its deregulation in cancer. J Cell Physiol 183(1):10–17. doi:10.1002/(SICI)1097-4652(200004)183:1<10::AID-JCP2>3.0.CO;2-I

    Article  CAS  PubMed  Google Scholar 

  29. Yue X, Zhang Z, Liang X, Gao L, Zhang X, Zhao D, Liu X, Ma H, Guo M, Spear BT, Gong Y, Ma C (2012) Zinc fingers and homeoboxes 2 inhibits hepatocellular carcinoma cell proliferation and represses expression of cyclins a and E. Gastroenterology 142(7):1559–1570 . doi:10.1053/j.gastro.2012.02.049e1552

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Youssef J, Badr M (2011) Peroxisome proliferator-activated receptors and cancer: challenges and opportunities. Br J Pharmacol 164(1):68–82. doi:10.1111/j.1476-5381.2011.01383.x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Hasmall SC, James NH, Macdonald N, Gonzalez FJ, Peters JM, Roberts RA (2000) Suppression of mouse hepatocyte apoptosis by peroxisome proliferators: role of PPARalpha and TNFalpha. Mutat Res 448(2):193–200

    Article  CAS  PubMed  Google Scholar 

  32. Gonzalez FJ, Peters JM, Cattley RC (1998) Mechanism of action of the nongenotoxic peroxisome proliferators: role of the peroxisome proliferator-activator receptor alpha. J Natl Cancer Inst 90(22):1702–1709

    Article  CAS  PubMed  Google Scholar 

  33. Nishimura J, Dewa Y, Muguruma M, Kuroiwa Y, Yasuno H, Shima T, Jin M, Takahashi M, Umemura T, Mitsumori K (2007) Effect of fenofibrate on oxidative DNA damage and on gene expression related to cell proliferation and apoptosis in rats. Toxicological Sciences: an Official Journal of the Society of Toxicology 97(1):44–54. doi:10.1093/toxsci/kfm011

    Article  CAS  Google Scholar 

  34. Shah YM, Morimura K, Yang Q, Tanabe T, Takagi M, Gonzalez FJ (2007) Peroxisome proliferator-activated receptor alpha regulates a microRNA-mediated signaling cascade responsible for hepatocellular proliferation. Mol Cell Biol 27(12):4238–4247. doi:10.1128/MCB.00317-07

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Fidaleo M (2009) Human health risk assessment for peroxisome proliferators: more than 30 years of research. Experimental and Toxicologic Pathology: Official Journal of the Gesellschaft fur Toxikologische Pathologie 61(3):215–221. doi:10.1016/j.etp.2008.09.002

    Article  Google Scholar 

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Authors and Affiliations

  1. Department of Healthcare Administration, Asia University, Taichung, 413, Taiwan

    Shu Nu Chang-Lee

  2. Division of Colorectal Surgery, Mackay Memorial Hospital, Taipei, 104, Taiwan

    Hsi-Hsien Hsu

  3. Mackay Medicine, Nursing and Management College, Taipei, 11260, Taiwan

    Hsi-Hsien Hsu

  4. Graduate Institute of Basic Medical Science, China Medical University, Taichung, 404, Taiwan

    Marthandam Asokan Shibu, Chih-Hao Tsai & Chih-Yang Huang

  5. Graduate Institute of Chinese Medicine, China Medical University, Taichung, 404, Taiwan

    Tsung-Jung Ho & Chih-Yang Huang

  6. Chinese Medicine Department, China Medical University Beigang Hospital, Yunlin, 651, Taiwan

    Tsung-Jung Ho

  7. Division of Colorectal Surgery, Taichung Veterans General Hospital, Taichung, 407, Taiwan

    Ming-Cheng Chen

  8. Division of Chest Medicine, Department of Internal Medicine, Taichung Armed Force General Hospital, Taichung, 411, Taiwan

    Chuan-Chou Tu

  9. Department of Biotechnology, Bharathiar University, Coimbatore, 641046, India

    Vijaya Padma Viswanadha

  10. Department of Biological Science and Technology, China Medical University, Taichung, 402, Taiwan

    Wei- Wen Kuo

  11. Department of Health and Nutrition Biotechnology, Asia University, Taichung, 413, Taiwan

    Chih-Yang Huang

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  1. Shu Nu Chang-Lee
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Correspondence to Chih-Yang Huang.

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This study is supported in part by Taiwan Ministry of Health and Welfare Clinical Trial and Research Center of Excellence (MOHW105-TDU-B-212-133,019).

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Chang-Lee, S.N., Hsu, HH., Shibu, M.A. et al. E2/ERβ Inhibits PPARα to Regulate Cell-Proliferation and Enhance Apoptosis in Hep3B-Hepatocellular Carcinoma. Pathol. Oncol. Res. 23, 477–485 (2017). https://doi.org/10.1007/s12253-016-0136-8

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  • DOI: https://doi.org/10.1007/s12253-016-0136-8

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