Estrogen deficiency reversibly induces telomere shortening in mouse granulosa cells and ovarian aging in vivo

Abstract

Estrogen is implicated as playing an important role in aging and tumorigenesis of estrogen responsive tissues; however the mechanisms underlying the mitogenic actions of estrogen are not fully understood. Here we report that estrogen deficiency in mice caused by tar-geted disruption of the aromatase gene results in a signi-ficant inhibition of telomerase maintenance of telomeres in mouse ovaries in a tissue-specific manner. The inhibition entails a significant shortening of telomeres and compromised proliferation in the follicular granulosa cell compartment of ovary. Gene expression analysis showed decreased levels of proto-oncogene c-Myc and the telomerase catalytic subunit, telomerase reverse transcriptase (TERT), in response to estrogen deficiency. Estrogen replacement therapy led to increases in TERT gene expression, telomerase activity, telomere length and ovarian tissue growth, thereby reinstating ovary development to normal in four weeks. Our data demonstrate for the first time that telomere maintenance is the primary mechanism mediating the mitogenic effect of estrogen on ovarian granulosa cell proliferation by up-regulating the genes of c-Myc and TERT in vivo. Estrogen deficiency or over-activity may cause ovarian tissue aging or tumorigenesis, respectively, through estrogen regulation of telomere remodeling.

Keywords

estrogen / telomerase reverse transcriptase (TERT) / telomeres / ovary / mice

References

[1] Baykal, A., Thompson, J.A., Xu, X.C., Hahn, W.C., Deavers, M.T., Malpica, A., Gershenson, D.M., Silva, E.G., and Liu, J. (2004). In situ human telomerase reverse transcriptase expression pattern in normal and neoplastic ovarian tissues. Oncol Rep 11 , 297-302 .
[2] Bayne, S., Jones, M.E., Li, H., and Liu, J.P. (2007). Potential roles for estrogen regulation of telomerase activity in aging. Ann N Y Acad Sci 1114, 48-55 .
[3] Bayne, S., Jones, M.E., Li, H., Pinto, A.R., Simpson, E.R., and Liu, J.P. (2008). Estrogen deficiency leads to telomerase inhibition, telomere shortening and reduced cell proliferation in the adrenal gland of mice. Cell Res 18, 1141-1150 .18936784
[4] Bj?rnstr?m, L., and Sj?berg, M. (2005). Mechanisms of estrogen receptor signaling: convergence of genomic and nongenomic actions on target genes. Mol Endocrinol 19, 833-842 .15695368
[5] Blasco, M.A. (2007). Telomere length, stem cells and aging. Nat Chem Biol 3, 640-649 .17876321
[6] Britt, K.L., Drummond, A.E., Dyson, M., Wreford, N.G., Jones, M.E., Simpson, E.R., and Findlay, J.K. (2001). The ovarian phenotype of the aromatase knockout (ArKO) mouse. J Steroid Biochem Mol Biol 79, 181-185 .11850223
[7] Britt, K.L., and Findlay, J.K. (2003). Regulation of the phenotype of ovarian somatic cells by estrogen. Mol Cell Endocrinol 202, 11-17 .12770724
[8] Britt, K.L., Kerr, J., O’Donnell, L., Jones, M.E., Drummond, A.E., Davis, S.R., Simpson, E.R., and Findlay, J.K. (2002). Estrogen regulates development of the somatic cell phenotype in the eutherian ovary. FASEB J 16, 1389-1397 .12205030
[9] Bullock, L.P., and Bardin, C.W. (1975). The prosence of estrogen receptor in kidneys from normal and androgen-insensitive tfm/y mice. Endocrinology 97, 1106-1111 .171143
[10] Chadeneau, C., Siegel, P., Harley, C.B., Muller, W.J., and Bacchetti, S. (1995). Telomerase activity in normal and malignant murine tissues. Oncogene 11, 893-898 .7675448
[11] Ciana, P., Raviscioni, M., Mussi, P., Vegeto, E., Que, I., Parker, M.G., Lowik, C., and Maggi, A. (2003). In vivo imaging of transcriptionally active estrogen receptors. Nat Med 9, 82-86 .12483206
[12] Filardo, E., Quinn, J., Pang, Y., Graeber, C., Shaw, S., Dong, J., and Thomas, P. (2007). Activation of the novel estrogen receptor G protein-coupled receptor 30 (GPR30) at the plasma membrane. Endocrinology 148, 3236-3245 .17379646
[13] Fisher, C.R., Graves, K.H., Parlow, A.F., and Simpson, E.R. (1998). Characterization of mice deficient in aromatase (ArKO) because of targeted disruption of the cyp19 gene. Proc Natl Acad Sci U S A 95, 6965-6970 .9618522
[14] Flores, I., and Blasco, M.A. (2009). A p53-dependent response limits epidermal stem cell functionality and organismal size in mice with short telomeres. PLoS One 4, e4934.19295915
[15] Flores, I., Cayuela, M.L., and Blasco, M.A. (2005). Effects of telomerase and telomere length on epidermal stem cell behavior. Science 309, 1253-1256 .16037417
[16] Fox, E.M., Andrade, J., and Shupnik, M.A. (2009). Novel actions of estrogen to promote proliferation: integration of cytoplasmic and nuclear pathways. Steroids 74, 622-627 .18996136
[17] Frasor, J., Danes, J.M., Komm, B., Chang, K.C., Lyttle, C.R., and Katzenellenbogen, B.S. (2003). Profiling of estrogen up- and down-regulated gene expression in human breast cancer cells: insights into gene networks and pathways underlying estrogenic control of proliferation and cell phenotype. Endocrinology 144, 4562-4574 .12959972
[18] Grasselli, A., Nanni, S., Colussi, C., Aiello, A., Benvenuti, V., Ragone, G., Moretti, F., Sacchi, A., Bacchetti, S., Gaetano, C., (2008). Estrogen receptor-alpha and endothelial nitric oxide synthase nuclear complex regulates transcription of human telomerase. Circ Res 103, 34-42 .18519947
[19] Greenberg, R.A., O’Hagan, R.C., Deng, H., Xiao, Q., Hann, S.R., Adams, R.R., Lichtsteiner, S., Chin, L., Morin, G.B., and DePinho, R.A. (1999). Telomerase reverse transcriptase gene is a direct target of c-Myc but is not functionally equivalent in cellular transformation. Oncogene 18, 1219-1226 .10022128
[20] Harrington, L., and Robinson, M.O. (2002). Telomere dysfunction: multiple paths to the same end. Oncogene 21, 592-597 .11850784
[21] Herrera, E., Samper, E., and Blasco, M.A. (1999). Telomere shortening in mTR-/- embryos is associated with failure to close the neural tube. EMBO J 18, 1172-1181 .10064584
[22] Hirshfield, A.N. (1991). Development of follicles in the mammalian ovary. Int Rev Cytol 124, 43-101 .2001918
[23] Jin, W., Chen, Y., Di, G.H., Miron, P., Hou, Y.F., Gao, H., and Shao, Z.M. (2008). Estrogen receptor (ER) beta or p53 attenuates ERalpha-mediated transcriptional activation on the BRCA2 promoter. J Biol Chem 283, 29671-29680 .18765668
[24] Jones, M.E., Thorburn, A.W., Britt, K.L., Hewitt, K.N., Wreford, N.G., Proietto, J., Oz, O.K., Leury, B.J., Robertson, K.M., Yao, S., (2000). Aromatase-deficient (ArKO) mice have a phenotype of increased adiposity. Proc Natl Acad Sci U S A 97, 12735-12740 .11070087
[25] Kanaya, T., Kyo, S., Hamada, K., Takakura, M., Kitagawa, Y., Harada, H., and Inoue, M. (2000). Adenoviral expression of p53 represses telomerase activity through down-regulation of human telomerase reverse transcriptase transcription. Clin Cancer Res 6, 1239-1247 .10778946
[26] Kyo, S., Takakura, M., Kanaya, T., Zhuo, W., Fujimoto, K., Nishio, Y., Orimo, A., and Inoue, M. (1999). Estrogen activates telomerase. Cancer Res 59, 5917-5921 .10606235
[27] Lacey, J.V. Jr, Mink, P.J., Lubin, J.H., Sherman, M.E., Troisi, R., Hartge, P., Schatzkin, A., and Schairer, C. (2002). Menopausal hormone replacement therapy and risk of ovarian cancer. JAMA 288, 334-341 .12117398
[28] Lavranos, T.C., Mathis, J.M., Latham, S.E., Kalionis, B., Shay, J.W., and Rodgers, R.J. (1999). Evidence for ovarian granulosa stem cells: telomerase activity and localization of the telomerase ribonucleic acid component in bovine ovarian follicles. Biol Reprod 61, 358-366 .10411512
[29] Lee, H.W., Blasco, M.A., Gottlieb, G.J., Horner, J.W. 2nd, Greider, C.W., and DePinho, R.A. (1998). Essential role of mouse telomerase in highly proliferative organs. Nature 392, 569-574 .9560153
[30] Ling, S., Zhou, L., Li, H., Dai, A., Liu, J.P., Komesaroff, P.A., and Sudhir, K. (2006). Effects of 17beta-estradiol on growth and apoptosis in human vascular endothelial cells: influence of mechanical strain and tumor necrosis factor-alpha. Steroids 71, 799-808 .16806337
[31] Liu, J.P., Chen, S.M., Cong, Y.S., Nicholls, C., Zhou, S.F., Tao, Z.Z., and Li, H. (2010). Regulation of telomerase activity by apparently opposing elements. Ageing Res Rev 9, 245-256 .20362078
[32] Liu, J.P., and Li, H. (2010). Telomerase in the ovary. Reproduction 140, 215-222 .20562297
[33] Meng, Y.G., Han, W.D., Zhao, Y.L., Huang, K., Si, Y.L., Wu, Z.Q., and Mu, Y.M. (2007). Induction of the LRP16 gene by estrogen promotes the invasive growth of Ishikawa human endometrial cancer cells through the downregulation of E-cadherin. Cell Res 17, 869-880 .17893710
[34] Miller, W.R., Bartlett, J.M., Canney, P., and Verrill, M. (2007a). Hormonal therapy for postmenopausal breast cancer: the science of sequencing. Breast Cancer Res Treat 103, 149-160 .17039263
[35] Miller, W.R., Larionov, A., Renshaw, L., Anderson, T.J., White, S., Hampton, G., Walker, J.R., Ho, S., Krause, A., Evans, D.B., (2007b). Aromatase inhibitors—gene discovery. J Steroid Biochem Mol Biol 106, 130-142 .17616392
[36] Misiti, S., Nanni, S., Fontemaggi, G., Cong, Y.S., Wen, J., Hirte, H.W., Piaggio, G., Sacchi, A., Pontecorvi, A., Bacchetti, S., (2000). Induction of hTERT expression and telomerase activity by estrogens in human ovary epithelium cells. [In Process Citation] Mol Cell Biol 20, 3764-3771 .10805720
[37] Payne, R.W., and Hellbaum, A.A. (1955). The effect of estrogens on the ovary of the hypophysectomized rat. Endocrinology 57, 193-199 .13251223
[38] Piontkewitz, Y., Sundfeldt, K., and Hedin, L. (1997). The expression of c-myc during follicular growth and luteal formation in the rat ovary in vivo. J Endocrinol 152, 395-406 .9071960
[39] Prowse, K.R., and Greider, C.W. (1995). Developmental and tissue-specific regulation of mouse telomerase and telomere length. Proc Natl Acad Sci U S A 92, 4818-4822 .7761406
[40] Revankar, C.M., Cimino, D.F., Sklar, L.A., Arterburn, J.B., and Prossnitz, E.R. (2005). A transmembrane intracellular estrogen receptor mediates rapid cell signaling. Science 307, 1625-1630 .15705806
[41] Robker, R.L., and Richards, J.S. (1998). Hormonal control of the cell cycle in ovarian cells: proliferation versus differentiation. Biol Reprod 59, 476-482 .9716543
[42] Rodgers, R.J., Irving-Rodgers, H.F., van Wezel, I.L., Krupa, M., and Lavranos, T.C. (2001). Dynamics of the membrana granulosa during expansion of the ovarian follicular antrum. Mol Cell Endocrinol 171, 41-48 .11165009
[43] Rufer, N. , Dragowska, W., Thornbury G., Roosnek E.,and Lansdop, P.M.,(1998). Telomere length dynamics in human lymphocyte subpopulations measured by flow cytometry. Nat Biotechnol 16 , 743-747
[44] Russo, V., Berardinelli, P., Martelli, A., Di Giacinto, O., Nardinocchi, D., Fantasia, D., and Barboni, B. (2006). Expression of telomerase reverse transcriptase subunit (TERT) and telomere sizing in pig ovarian follicles. J Histochem Cytochem 54, 443-455 .16400001
[45] Santos, G.F., Scott, G.K., Lee, W.M.F., Liu, E., and Benz, C. (1988). Estrogen-induced post-transcriptional modulation of c-myc proto-oncogene expression in human breast cancer cells. J Biol Chem 263, 9565-9568 .3290209
[46] Sarkar, P., Shiizaki, K., Yonemoto, J., and Sone, H. (2006). Activation of telomerase in BeWo cells by estrogen and 2,3,7,8-tetrachlorodibenzo-p-dioxin in co-operation with c-Myc. Int J Oncol 28, 43-51 .16327978
[47] Schoeftner, S., and Blasco, M.A. (2009). A 'higher order' of telomere regulation: telomere heterochromatin and telomeric RNAs. EMBO J 28 , 2323-2336 .
[48] Segars, J.H., and Driggers, P.H. (2002). Estrogen action and cytoplasmic signaling cascades. Part I: membrane-associated signaling complexes. Trends Endocrinol Metab 13, 349-354 .12217492
[49] Shats, I., Milyavsky, M., Tang, X., Stambolsky, P., Erez, N., Brosh, R., Kogan, I., Braunstein, I., Tzukerman, M., Ginsberg, D., (2004). p53-dependent down-regulation of telomerase is mediated by p21waf1. J Biol Chem 279, 50976-50985 .15371422
[50] Shay, J.W., and Bacchetti, S. (1997). A survey of telomerase activity in human cancer. Eur J Cancer 33, 787-791 .9282118
[51] Toda, K., Takeda, K., Okada, T., Akira, S., Saibara, T., Kaname, T., Yamamura, K., Onishi, S., and Shizuta, Y. (2001). Targeted disruption of the aromatase P450 gene (Cyp19) in mice and their ovarian and uterine responses to 17beta-oestradiol. J Endocrinol 170, 99-111 .11431142
[52] van der Burg, B., van Selm-Miltenburg, A.J., de Laat, S.W., and van Zoelen, E.J. (1989). Direct effects of estrogen on c-fos and c-myc protooncogene expression and cellular proliferation in human breast cancer cells. Mol Cell Endocrinol 64, 223-228 .2507374
[53] Yamagata, Y., Nakamura, Y., Umayahara, K., Harada, A., Takayama, H., Sugino, N., and Kato, H. (2002). Changes in telomerase activity in experimentally induced atretic follicles of immature rats. Endocr J 49, 589-595 .12625407