Cell Cycle Regulation in the Estrogen Receptor Beta (ESR2)-Overexpressing Hep3B Hepatocellular Carcinoma Cell Line

10.4077/CJP.2015.BAC239

Yi-Sheng Liu；Ying-Lan Tsai；Yu-Lan Yeh；Li-Chin Chung；Su-Ying Wen；Chia-Hua Kuo；Yueh-Min Lin；V. Vijaya Padma；V. Bharath Kumar；Chih-Yang Huang

c-myc ； cyclin D1 ； ESR2 ； Hep3B cells

The Chinese Journal of Physiology

58卷2期（2015 / 04 / 30）

134 - 140

英文

Epidemiological studies and experimental data have shown that the incidences of hepatocellular carcinoma in men are more frequent than in women. Evidence suggests that imbalance of hormones, including estrogen, androgen, prolactin, and growth hormone, modifies liver tumorigenesis. In this present study, we investigated how estrogen and estrogen receptor 2 (ESR2), regulates the cell cycle mechanism in Hep3B hepatocellular carcinoma cell line. Our results showed that ESR2 overexpression in the presence of 10^(-8) M 17-β-estradiol downregulated c-myc and cyclin D1 expression and simultaneously upregulated p27 expression. However, flow cytometry and MTT assays showed only minor G_1 phase arrest without affecting cell viability. Taken together, these observations indicate that ESR2 is required to lower tumorigenesis in males by altering cell cycle proteins in a ligand-dependent manner.

1. Fan, M.J.,Huang-Liu, R.,Shen, C.Y.,Ju, D.T.,Lin, Y.M.,Pai, P.,Huang, P.Y.,Ho, T.J.,Tsai, F.J.,Tsai, C.H.,Huang, C.Y.(2014).Reduction of TLR4 mRNA stability and protein expressions through inhibiting cytoplasmic translocation of HuR transcription factor by E2 and/or ERα in LPS-treated H9c2 cardiomyoblast cells.Chinese J. Physiol.,57,8-18.
   連結：
2. Liu, F.,Gao, X.,Yu, H.,Yuan, D.,Zhang, J.,He, Y.,Yue, L.(2013).Effects of expression of exogenous cyclin G1 on proliferation of human endometrial carcinoma cells.Chinese J. Physiol.,56,83-89.
   連結：
3. Bardin, A.,Hoffmann, P.,Boulle, N.,Katsaros, D.,Vignon, F.,Pujol, P.,Lazennec, G.(2004).Involvement of estrogen receptor β in ovarian carcinogenesis.Cancer Res.,64,5861-5869.
4. Di Maio, M.,De Maio, E.,Perrone, F.,Pignata, S.,Daniele, B.(2002).Hepatocellular carcinoma: systemic treatments.J. Clin. Gastroenterol.,35,S109-S114.
5. Doisneau-Sixou, S.F.,Sergio, C.M.,Carroll, J.S.,Hui, R.,Musgrove, E.A.,Sutherland, R.L.(2003).Estrogen and antiestrogen regulation of cell cycle progression in breast cancer cells.Endocr.-Relat. Cancer,10,179-186.
6. El-Serag, H.B.(2002).Hepatocellular carcinoma: an epidemiologic view.J. Clin. Gastroenterol.,35,S72-S78.
7. Foley, E.F.,Jazaeri, A.A.,Shupnik, M.A.,Jazaeri, O.,Rice, L.W.(2000).Selective loss of estrogen receptor β in malignant human colon.Cancer Res.,60,245-248.
8. Foster, J.S.,Fernando, R.I.,Ishida, N.,Nakayama, K.I.,Wimalasena, J.(2003).Estrogens down-regulate p27Kip1 in breast cancer cells through Skp2 and through nuclear export mediated by the ERK pathway.J. Biol. Chem.,278,41355-41366.
9. Foster, J.S.,Henley, D.C.,Ahamed, S.,Wimalasena, J.(2001).Estrogens and cell-cycle regulation in breast cancer.Trends Endocrinol. Metab.,12,320-327.
10. Fuqua, S.A.W.,Schiff, R.,Parra, I.,Moore, J.T.,Mohsin, S.K.,Osborne, C.K.,Clark, G.M.,Allred, D.C.(2003).Estrogen receptor β protein in human breast cancer: correlation with clinical tumor parameters.Cancer Res.,63,2434-2439.
11. Hall, J.M.,Couse, J.F.,Korach, K.S.(2001).The multifaceted mechanisms of estradiol and estrogen receptor signaling.J. Biol. Chem.,276,36869-36872.
12. Hartman, J.,Lindberg, K.,Morani, A.,Inzunza, J.,Strom, A.,Gustafsson, J.A.(2006).Estrogen receptor β inhibits angiogenesis and growth of T47D breast cancer xenografts.Cancer Res.,66,11207-11213.
13. Hayashi, S.I.,Eguchi, H.,Tanimoto, K.,Yoshida, T.,Omoto, Y.,Inoue, A.,Yoshida, N.,Yamaguchi, Y.(2003).The expression and function of estrogen receptor α and β in human breast cancer and its clinical application.Endocr.-Relat. Cancer,10,193-202.
14. Iavarone, M.,Lampertico, P.,Seletti, C.,Francesca Donato, M.,Ronchi, G.,del Ninno, E.,Colombo, M.(2003).The clinical and pathogenetic significance of estrogen receptor-β expression in chronic liver diseases and liver carcinoma.Cancer,98,529-534.
15. Kalra, M.,Mayes, J.,Assefa, S.,Kaul, A.K.,Kaul, R.(2008).Role of sex steroid receptors in pathobiology of hepatocellular carcinoma.World J. Gastroenterol.,14,5945-5961.
16. Kuiper, G.G.,Carlsson, B.,Grandien, K.,Enmark, E.,Haggblad, J.,Nilsson, S.,Gustafsson, J.A.(1997).Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors α and β.Endocrinology,138,863-870.
17. Lazennec, G.,Bresson, D.,Lucas, A.,Chauveau, C.,Vignon, F.(2001).ER beta inhibits proliferation and invasion of breast cancer cells.Endocrinology,142,4120-4130.
18. Lin, Y.M.,Velmurugan, B.K.,Yeh, Y.L.,Tu, C.C.,Ho, T.J.,Lai, T.Y.,Tsai, C.H.,Tsai, F.J.,Tsai, C.H.,Huang, C.Y.(2013).Activation of estrogen receptors with E2 downregulates peroxisome proliferator-activated receptor γ in hepatocellular carcinoma.Oncol. Rep.,30,3027-3031.
19. Mishkin, S.Y.,Farber, E.,Ho, R.K.,Mulay, S.,Mishkin, S.(1983).Evidence for the hormone dependency of hepatic hyperplastic nodules: inhibition of malignant transformation after exogenous 17β-estradiol and tamoxifen.Hepatology,3,308-316.
20. Nakatani, T.,Roy, G.,Fujimoto, N.,Asahara, T.,Ito, A.(2001).Sex hormone dependency of diethylnitrosamine-induced liver tumors in mice and chemoprevention by leuprorelin.Jpn. J. Cancer Res.,92,249-256.
21. Newcomb, P.A.,Zheng, Y.Y.,Chia, V.M.,Morimoto, L.M.,Doria-Rose, V.P.,Templeton, A.,Thibodeau, S.N.,Potter, J.D.(2007).Estrogen plus progestin use, microsatellite instability, and the risk of colorectal cancer in women.Cancer Res.,67,7534-7539.
22. Paruthiyil, S.,Parmar, H.,Kerekatte, V.,Cunha, G.R.,Firestone, G.L.,Leitman, D.C.(2004).Estrogen receptor β inhibits human breast cancer cell proliferation and tumor formation by causing a G2 cell cycle arrest.Cancer Res.,64,423-428.
23. Pisani, P.,Parkin, D.M.,Bray, F.,Ferlay, J.(1999).Estimates of the worldwide mortality from 25 cancers in 1990.Int. J. Cancer,83,18-29.
24. Sert, F.,Ozsaran, Z.,Eser, E.,Alanyali, S.D.,Haydaroglu, A.,Aras, A.(2013).Quality of life assessment in women with breast cancer: a prospective study including hormonal therapy.J. Breast Cancer,16,220-228.
25. Shimizu, I.,Yasuda, M.,Mizobuchi, Y.,Ma, Y.R.,Liu, F.,Shiba, M.,Horie, T.,Ito, S.(1998).Suppressive effect of oestradiol on chemical hepatocarcinogenesis in rats.Gut,42,112-119.
26. Strom, A.,Hartman, J.,Foster, J.S.,Kietz, S.,Wimalasena, J.,Gustafsson, J.A.(2004).Estrogen receptor β inhibits 17β-estradiolstimulated proliferation of the breast cancer cell line T47D.Proc. Natl. Acad. Sci. USA,101,1566-1571.
27. Tsutsui, S.,Yamamoto, R.,Iishi, H.,Tatsuta, M.,Tsuji, M.,Terada, N.(1992).Promoting effect of ovariectomy on hepatocellular tumorigenesis induced in mice by 3'-methyl-4-dimethylaminoazobenzene.Virchows Arch., B, Cell Pathol.,62,371-375.
28. Wang, A.G.,Lee, K.Y.,Kim, S.Y.,Choi, J.Y.,Lee, K.H.,Kim, W.H.,Wang, H.J.,Kim, J.M.,Park, M.G.,Yeom, Y.I.,Kim, N.S.,Yu, D.Y.,Lee, D.S.(2006).The expression of estrogen receptors in hepatocellular carcinoma in Korean patients.Yonsei Med. J.,47,811-816.
29. Weihua, Z.,Makela, S.,Andersson, L.C.,Salmi, S.,Saji, S.,Webster, J.I.,Jensen, E.V.,Nilsson, S.,Warner, M.,Gustafsson, J.A.(2001).A role for estrogen receptor β in the regulation of growth of the ventral prostate.Proc. Natl. Acad. Sci. USA,98,6330-6335.
30. Yeh, S.H.,Chen, P.J.(2010).Gender disparity of hepatocellular carcinoma: the roles of sex hormones.Oncology,78(Suppl 1),172-179.