An oestrogen receptor β-selective agonist exerts anti-neoplastic effects in experimental intrahepatic cholangiocarcinoma

      Abstract

      Background

      Cholangiocarcinoma cells over-express oestrogen receptor-β, which displays anti-proliferative and pro-apoptotic effects.

      Aim

      To evaluate the effects of a newly developed and highly selective oestrogen receptor-β agonist (KB9520) on experimental intrahepatic cholangiocarcinoma.

      Methods

      In vitro, the effects of KB9520 on apoptosis and proliferation of HuH-28 cells, HuH-28 cells with selective oestrogen receptor-β silencing (by small interfering RNA), HepG2 cells (oestrogen receptor-α and oestrogen receptor-β negative) and HepER3 cells (HepG2 cells transformed to stably express oestrogen receptor-α) were evaluated. In vivo, the effects of KB9520 on experimental intrahepatic cholangiocarcinoma, induced by thioacetamide administration were tested.

      Results

      In vitro, KB9520 induced apoptosis and inhibited proliferation of HuH-28 cells. KB9520 effects were absent in cells lacking oestrogen receptor-α and β (HepG2) and in cells expressing only oestrogen receptor-α (HepER3); its pro-apoptotic effect was impaired in cells where oestrogen receptor-β expression was decreased by specific small interfering RNA. In vivo, KB9520 inhibited experimental intrahepatic cholangiocarcinoma development in thioacetamide-treated rats and promoted tumour regression in rats where tumour was already established. In treated animals, tumour areas showed reduced proliferation but increased apoptosis.

      Conclusions

      KB9520 induced apoptosis in cholangiocarcinoma by selectively acting on oestrogen receptor-β, suggesting that oestrogen receptor-β selective agonists may be a novel and effective therapeutic option for the medical treatment of intrahepatic cholangiocarcinoma.

      Keywords

      To read this article in full you will need to make a payment

      References

        • Alvaro D.
        • Cannizzaro R.
        • Labianca R.
        • et al.
        Cholangiocarcinoma: a position paper by the Italian Society of Gastroenterology (SIGE), the Italian Association of Hospital Gastroenterology (AIGO), the Italian Association of Medical Oncology (AIOM) and the Italian Association of Oncological Radiotherapy (AIRO).
        Dig Liver Dis. 2010; 42: 831-838
        • Blechacz B.
        • Gores G.J.
        Cholangiocarcinoma: advances in pathogenesis, diagnosis, and treatment.
        Hepatology. 2008; 48: 308-321
        • Khan S.A.
        • Thomas H.C.
        • Davidson B.R.
        • et al.
        Cholangiocarcinoma.
        Lancet. 2005; 366: 1303-1314
        • Patel T.
        Worldwide trends in mortality from biliary tract malignancies.
        BMC Cancer. 2002; 2: 10
        • Alvaro D.
        • Barbaro B.
        • Franchitto A.
        • et al.
        Estrogens and insulin-like growth factor 1 modulate neoplastic cell growth in human cholangiocarcinoma.
        Am J Pathol. 2006; 169: 877-888
        • Mancino A.
        • Mancino M.G.
        • Glaser S.S.
        • et al.
        Estrogens stimulate the proliferation of human cholangiocarcinoma by inducing the expression and secretion of vascular endothelial growth factor.
        Dig Liver Dis. 2009; 41: 156-163
        • Alvaro D.
        • Mancino M.G.
        • Onori P.
        • et al.
        Estrogens and the pathophysiology of the biliary tree.
        World J Gastroenterol. 2006; 12: 3537-3545
        • Cheng G.
        • Li Y.
        • Omoto Y.
        • et al.
        Differential regulation of estrogen receptor (ER)alpha and ERbeta in primate mammary gland.
        J Clin Endocrinol Metab. 2005; 90: 435-444
        • Imamov O.
        • Morani A.
        • Shim G.J.
        • et al.
        Estrogen receptor beta regulates epithelial cellular differentiation in the mouse ventral prostate.
        Proc Natl Acad Sci U S A. 2004; 101: 9375-9380
        • Morani A.
        • Barros R.P.
        • Imamov O.
        • et al.
        Lung dysfunction causes systemic hypoxia in estrogen receptor beta knockout (ERbeta−/−) mice.
        Proc Natl Acad Sci USA. 2006; 103: 7165-7169
        • Wada-Hiraike O.
        • Imamov O.
        • Hiraike H.
        • et al.
        Role of estrogen receptor beta in colonic epithelium.
        Proc Natl Acad Sci USA. 2006; 103: 2959-2964
        • Warner M.
        • Gustafsson J.A.
        The role of estrogen receptor beta (ERbeta) in malignant diseases—a new potential target for antiproliferative drugs in prevention and treatment of cancer.
        Biochem Biophys Res Commun. 2010; 396: 63-66
      1. Yakimchuk K, Iravani M, Sharif Hasni M, et al. Effect of ligand-activated estrogen receptor β on lymphoma growth in vitro and in vivo. Leukemia; In press, https://doi.org/10.1038/leu.2011.68.

        • Barkhem T.
        • Andersson-Ross C.
        • Hoglund M.
        • et al.
        Characterization of the “estrogenicity” of tamoxifen and raloxifene in HepG2 cells: regulation of gene expression from an ERE controlled reporter vector versus regulation of the endogenous SHBG and PS2 genes.
        J Steroid Biochem Mol Biol. 1997; 62: 53-64
        • Franchitto A.
        • Torrice A.
        • Semeraro R.
        • et al.
        Prostate apoptosis response-4 is expressed in normal cholangiocytes, is down-regulated in human cholangiocarcinoma, and promotes apoptosis of neoplastic cholangiocytes when induced pharmacologically.
        Am J Pathol. 2010; 177: 1779-1790
        • Fava G.
        • Alpini G.
        • Rychlicki C.
        • et al.
        Leptin enhances cholangiocarcinoma cell growth.
        Cancer Res. 2008; 68: 6752-6761
        • Alvaro D.
        • Alpini G.
        • Onori P.
        • et al.
        Estrogens stimulate proliferation of intrahepatic biliary epithelium in rats.
        Gastroenterology. 2000; 119: 1681-1691
        • Marzioni M.
        • Francis H.
        • Benedetti A.
        • et al.
        Ca2+-dependent cytoprotective effects of ursodeoxycholic and tauroursodeoxycholic acid on the biliary epithelium in a rat model of cholestasis and loss of bile ducts.
        Am J Pathol. 2006; 168: 398-409
        • Marzioni M.
        • Alpini G.
        • Saccomanno S.
        • et al.
        Exendin-4, a glucagon-like peptide 1 receptor agonist, protects cholangiocytes from apoptosis.
        Gut. 2009; 58: 990-997
        • De Minicis S.
        • Candelaresi C.
        • Marzioni M.
        • et al.
        Role of endogenous opioids in modulating HSC activity in vitro and liver fibrosis in vivo.
        Gut. 2008; 57: 352-364
        • Isse K.
        • Specht S.M.
        • Lunz 3rd, J.G.
        • et al.
        Estrogen stimulates female biliary epithelial cell interleukin-6 expression in mice and humans.
        Hepatology. 2010; 51: 869-880
        • Sampson L.K.
        • Vickers S.M.
        • Ying W.
        • et al.
        Tamoxifen-mediated growth inhibition of human cholangiocarcinoma.
        Cancer Res. 1997; 57: 1743-1749
        • Heldring N.
        • Pike A.
        • Andersson S.
        • et al.
        Estrogen receptors: how do they signal and what are their targets.
        Physiol Rev. 2007; 87: 905-931
        • Ogawa S.
        • Inoue S.
        • Watanabe T.
        • et al.
        Molecular cloning and characterization of human estrogen receptor betacx: a potential inhibitor ofestrogen action in human.
        Nucleic Acids Res. 1998; 26: 3505-3512
        • Blizzard T.A.
        • Selective estrogen receptor modulator medicinal chemistry at Merck. A review
        Curr Top Med Chem. 2008; 8: 792-812
        • Harris H.A.
        • Albert L.M.
        • Leathurby Y.
        • et al.
        Evaluation of an estrogen receptor-beta agonist in animal models of human disease.
        Endocrinology. 2003; 144: 4241-4249
        • Shim G.J.
        • Wang L.
        • Andersson S.
        • et al.
        Disruption of the estrogen receptor beta gene in mice causes myeloproliferative disease resembling chronic myeloid leukemia with lymphoid blast crisis.
        Proc Natl Acad Sci USA. 2003; 100: 6694-6699
        • Pinton G.
        • Thomas W.
        • Bellini P.
        • et al.
        Estrogen receptor β exerts tumor repressive functions in human malignant pleural mesotheliom via EGFR inactivation and affects response to gefitinib.
        PLoS ONE. 2010; 5: e14110
        • Suzuki F.
        • Akahira J.
        • Miura I.
        • et al.
        Loss of estrogen receptor beta isoform expression and its correlation with aberrant DNA methylation of the 5’-untranslated region in human epithelial ovarian carcinoma.
        Cancer Sci. 2008; 99: 2365-2372
        • Hartman J.
        • Lindberg K.
        • Morani A.
        • et al.
        Estrogen receptor beta inhibits angiogenesis and growth of T47D breast cancer xenografts.
        Cancer Res. 2006; 66: 11207-11213
        • Dondi D.
        • Piccolella M.
        • Biserni A.
        • et al.
        Estrogen receptor beta and the progression of prostate cancer: role of 5alpha-androstane-3beta, 17beta-diol.
        Endocr Relat Cancer. 2010; 17: 731-742
        • Barkhem T.
        • Carlsson B.
        • Nilsson Y.
        • et al.
        Differential response of estrogen receptor alpha and estrogen receptor beta to partial estrogen agonists/antagonists.
        Mol Pharmcol. 1998; 54: 105-112
        • Kohigashi K.
        • Fukuda Y.
        • Imura H.
        Estrogen receptors in hepatocellular carcinoma: is endocrine therapy for hepatocellular carcinoma likely to be effective?.
        Gastroenterol Jpn. 1987; 22: 322-330
        • Ohnishi S.
        • Murakami T.
        • Moriyama T.
        • et al.
        Androgen and estrogen receptors in hepatocellular carcinoma and in the surrounding noncancerous liver tissue.
        Hepatology. 1986; 6: 440-443
        • Eagon P.K.
        • Francavilla A.
        • DiLeo A.
        • et al.
        Quantitation of estrogen and androgen receptors in hepatocellular carcinoma and adjacent normal human liver.
        Dig Dis Sci. 1991; 36: 1303-1308
        • Iavarone M.
        • Lampertico P.
        • Seletti C.
        • et al.
        The clinical and pathogenetic significance of estrogen receptor-beta expression in chronic liver diseases and liver carcinoma.
        Cancer. 2003; 98: 529-534
        • Kalra M.
        • Mayes J.
        • Assefa S.
        • et al.
        Role of sex steroid receptors in pathobiology of hepatocellular carcinoma.
        World J Gastroenterol. 2008; 14: 5945-5961