Estrogen Receptor-α Mediates the Effects of Estradiol on Telomerase Activity in Human Mesenchymal Stem Cells

  • Cha, Young (College of Medicine, Pochon CHA University) ;
  • Kwon, Su Jin (College of Medicine, Pochon CHA University) ;
  • Seol, Wongi (Institute for Brain Science and Technology, Inje University) ;
  • Park, Kyung-Soon (College of Medicine, Pochon CHA University)
  • Received : 2008.04.14
  • Accepted : 2008.08.22
  • Published : 2008.11.30

Abstract

Sex steroid hormone receptors play a central role in modulating telomerase activity, especially in cancer cells. However, information on the regulation of steroid hormone receptors and their distinct functions on telomerase activity within the mesenchymal stem cell are largely unavailable due to low telomerase activity in the cell. In this study, the effects of estrogen ($E_2$) treatment and function of estrogen receptor alpha ($ER{\alpha}$) and estrogen receptor beta ($ER{\beta}$) on telomerase activity were investigated in human mesenchymal stem cells (hMSCs). Telomerase activity and mRNA expression of the catalytic subunit of telomerase (hTERT) were upregulated by treatment of the cells with $E_2$. The protein concentration of $ER{\alpha}$ was also increased by $E_2$ treatment, and enhancement of $ER{\alpha}$ accumulation in the nucleus was clearly detected with immunocytochemistry. When $ER{\alpha}$ expression was reduced by siRNA transfection into hMSCs, the effect of $E_2$ on the induction of hTERT expression and telomerase activity was diminished. In contrast, the transient overexpression of $ER{\alpha}$ increased the effect of $E_2$ on the expression of hTERT mRNA. These findings indicate that the activation of hTERT expression and telomerase activity by $E_2$ in hMSCs depends on $ER{\alpha}$, but not on $ER{\beta}$.

Keywords

AKT1;estrogen receptor;mesenchymal stem cells;telomerase

Acknowledgement

Supported by : Korea Science and Engineering Foundation

References

  1. Harley, C.B. (1995). Telomeres and telomerase in aging and cancer. Curr. Opin. Genet. Dev. 5, 249-255 https://doi.org/10.1016/0959-437X(95)80016-6
  2. Hoelters, J., Ciccarella, M., Drechsel, M., Geissler, C., Gulkan, H., Bocker, W., Schieker, M., Jochum, M., and Neth, P. (2005). Nonviral genetic modification mediates effective transgene expression and functional RNA interference in human mesenchymal stem cells. J. Gene Med. 7, 718-728 https://doi.org/10.1002/jgm.731
  3. Kyo, S., Kanaya, T., Takakura, M., Tanaka, M., and Inoue, M. (1999a). Human telomerase reverse transcriptase as a critical determinant of telomerase activity in normal and malignant endometrial tissues. Int. J. Cancer 80, 60-63 https://doi.org/10.1002/(SICI)1097-0215(19990105)80:1<60::AID-IJC12>3.0.CO;2-E
  4. Paech, K., Webb, P., Kuiper, G.G.J.M., Nilsson, S., Gustafsson, J.A., Kushner, P.J., and Scanlan, T.S. (1997). Differential ligand activation of estrogen receptors ER$\alpha$ and ER$\beta$ at AP1 sites. Science 277, 1508-1510 https://doi.org/10.1126/science.277.5331.1508
  5. Richard, D.J., Subramaniam, M., and Spelsberg, T.C. (1999). Molecular and cellular mechanisms of estrogen action on the skeleton. J. Cell. Biochem. 75, 123-132 https://doi.org/10.1002/(SICI)1097-4644(1999)75:32+<123::AID-JCB15>3.0.CO;2-K
  6. Takakura, M., Kyo, S., Kanaya, T., Tanaka, M., and Inoue, M. (1998). Expression of human telomerase subunits and correlation with telomerase activity in cervical cancer. Cancer Res. 58, 1558-1561
  7. Kimura, A., Ohmichi, M., Kawagoe, J., Kyo, S., Mabuchi, S., Takahashi, T., Ohshima, C., Aromoto-Ishida, E., Nishio, Y., Inoue, M., et al. (2004). Induction of hTERT expression and phosphorylation by estrogen via Akt cascade in human ovarian cancer cell lines. Oncogene 23, 4505-4515 https://doi.org/10.1038/sj.onc.1207582
  8. Ahn, S.E., Kim, S., Park, K.H., Moon, S.H., Lee, H.J., Kim, G.J., Lee, Y.J., Park, K.H., Cha, K.Y., and Chung, H.M. (2006). Primary bone-derived cells induce osteogenic differentiation without exogenous factors in human embryonic stem cells. Biochem. Biophys. Res. Commun. 340, 403-408 https://doi.org/10.1016/j.bbrc.2005.12.020
  9. Misiti, S., Nanni, S., Fontemaggi, G., Cong, Y.S., Wen, J., Hirte, H.W., Piaggio, G., Sacchi, A., Pontecorvi, A., Bacchetti, S., et al. (2000). Induction of hTERT expression and telomerase activity by estrogens in human ovary epithelium cells. Mol. Cell. Biol. 20, 3764-3771 https://doi.org/10.1128/MCB.20.11.3764-3771.2000
  10. Wege, H., Chui, M.S., Le, H.T., Tran, J.M., and Zern, M.A. (2003). SYBR Green real-time telomeric repeat amplification protocol for the rapid quantification of telomerase activity. Nucleic Acids Res. 31, e3 https://doi.org/10.1093/nar/gng003
  11. Zimmermann, S., Voss, M., Kaiser, S., Kapp, U., Waller, C.F., and Martens, U.M. (2003). Lack of telomerase activity in human mesenchymal stem cells. Leukemia 17, 1146-1149 https://doi.org/10.1038/sj.leu.2402962
  12. Beato, M., Herrlich, P., and Schutz, G. (1995). Steroid hormone receptors: many actors in search of a plot. Cell 83, 851-857 https://doi.org/10.1016/0092-8674(95)90201-5
  13. Greider, C.W., and Blackburn, E.H. (1985). Identification of a specific telomere terminal transferase activity in Tetrahymena extracts. Cell 43, 405-413 https://doi.org/10.1016/0092-8674(85)90170-9
  14. Kim, N.W., and We, F. (1997). Advances in quantification and characterization of telomerase activity by the telomeric repeat amplification protocol (TRAP). Nucleic Acids Res. 25, 2595-2597 https://doi.org/10.1093/nar/25.13.2595
  15. Ascenzi, P., Bocedi, A., and Marino, M. (2006). Structure-function relationships of estrogen receptor-$\alpha$ and $\beta$: Impact on human health. Mol. Aspects Med. 27, 299-402 https://doi.org/10.1016/j.mam.2006.07.001
  16. Seol, W., Hanstein, B., Brown, M., and Moore, D.D. (1998). Inhibition of estrogen receptor action by the orphan receptor SHP (short heterodimer partner). Mol. Endocrinol. 12, 1551-1557 https://doi.org/10.1210/me.12.10.1551
  17. Doshida, M., Ohmichi, M., Tsutsumi, S., Kawagoe, J., Takahashi, T., Du, B., Mori-Abe, A., Ohta, T., Saitoh-Sekiguchi, M., Takahashi, K., et al. (2006). Raloxifene increases proliferation and upregulates telomerase activity in human umbilical vein endothelial cells. J. Biol. Chem. 281, 24270-24278 https://doi.org/10.1074/jbc.M513251200
  18. Greider, C.W., and Blackburn, E.H. (1989). A telomeric sequence in the RNA of Tetrahymena telomerase required for telomere repeat synthesis. Nature 337, 331-337 https://doi.org/10.1038/337331a0
  19. Hall, J.M., Couse, J.F., and Korach, K.S. (2001). The multifaceted mechanisms of estradiol and estrogen receptor signaling. J. Biol. Chem. 276, 36869-36872 https://doi.org/10.1074/jbc.R100029200
  20. Klinge, C.M., Blankenship, K.A., Risinger, K.E., Bhatnagar, S., Noisin, E.L., Sumanasekera, W.K., Zhao, L., Brey, D.M., and Keynton, R.S. (2005). Resveratrol and estradiol rapidly activate MAPK signaling through estrogen receptors $\alpha$ and $\beta$ in endothelial cells. J. Biol. Chem. 280, 7460-7468 https://doi.org/10.1074/jbc.M411565200
  21. Kyo, S., Kanaya, T., Takakura, M., Tanaka, M., Yamashita, A., Inoue, H., and Inoue, M. (1999b). Expression of human telomerase subunits in ovarian malignant, borderline and benign tumors. Int. J. Cancer 80, 804-809 https://doi.org/10.1002/(SICI)1097-0215(19990315)80:6<804::AID-IJC2>3.0.CO;2-B
  22. Engelhardt, M., Kumar, R., Albanell, J., Pettengell, R., Han, W., and Moore, M.A. (1997). Telomerase regulation, cell cycle, and telomerase stability in primitive hematopoietic cells. Blood 90, 182-193
  23. Harle-Bachor, C., and Boukamp, P. (1996). Telomerase activity in the regenerative basal layer of the epidermis in human skin and in immortal and carcinoma-derived skin keratinocytes. Proc. Natl. Acad. Sci. USA 93, 6476-6481
  24. Kolquist, K.A., Ellisen, L.W., Counter, C.M., Meyerson, M., Tan, L.K., Weinberg, R.A., Haber, D.A., and Gerald, W.L. (1998). Expression of TERT in early premalignant lesions and a subset of cells in normal tissues. Nat. Genet. 19, 182-186 https://doi.org/10.1038/554
  25. Parsch, D., Fellenberg, J., Brummendorf, T.H., Eschlbeck, A.M., and Richter, W. (2004). Telomerase length and telomerase activity during expansion and differentiation of human mesenchymal stem cells and chondrocytes. J. Mol. Med. 82, 49-55 https://doi.org/10.1007/s00109-003-0506-z