Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

Inhibitory Effects of High Concentrations of Estrogen, Progesterone and Tamoxifen on Proliferation of HeLa in Culture

배양된 HeLa 세포에서 고농도의 에스트로겐, 프로게스테론 및 타목시펜의 세포증식 억제효과

  • Min, Gye-Sik (Department of Pharmaceutical Engineering, Gyeongnam National University of Science & Technology)
  • 민계식 (경남과학기술대학교 제약공학과)
  • Received : 2011.10.31
  • Accepted : 2011.11.11
  • Published : 2011.12.31
Download PDF
⟨ Previous Next ⟩

Abstract

This study examined the effects of estrogen, progesterone and tamoxifen at different concentrations and treatment periods on proliferation of a human cervical carcinoma cell line, HeLa, in culture, based on MTT assay. Estrogen did not have an effect on the cellular proliferation in concentrations up to $1{\mu}g$/ml for treatment periods of between 2.5 and 6 days, but significantly inhibited proliferation at a higher concentration of $10{\mu}g$/ml in a progressive manner with increasing treatment periods. Also, treatment of HeLa with more than $10{\mu}g$/ml of progesterone for 2.5 days significantly inhibited proliferation and caused a concentration-dependent inhibition with 4 days of treatment. However, longer treatment with progesterone for 6 days abolished the concentration-dependent inhibitory effect on cellular proliferation observed with the 4-day treatment period. Furthermore, tamoxifen required a higher concentration ($100{\mu}g$/ml) than estrogen to bring about the inhibitory effect on the HeLa proliferation. These results suggest that high concentrations of estrogen, progesterone and tamoxifen may suppress proliferation of HeLa, and both the concentration and the treatment period may also influence their inhibitory effects on cellular proliferation.

이 연구는 에스트로겐, 프로게스테론 및 타목시펜의 각각 다른 농도와 처리기간에 따라 배양된 사람 난소유래암세포주인 HeLa 세포의 증식에 미치는 영향을 MTT 분석에 의해 조사하였다. 에스트로겐은 2.5~6일의 처리기간 동안 $1{\mu}g$/ml의 농도까지는 세포증식에 영향을 주지 않았지만, 더 높은 $10{\mu}g$/ml의 농도에서는 처리기간의 증가에 따라 점진적으로 현저하게 세포증식을 억제하였다. 또한 $10{\mu}g$/ml 농도 이상의 프로게스테론을 2.5일 동안 처리할 경우 HeLa 세포의 증식을 현저하게 억제하였으며, 4일 동안 처리시에는 농도-의존성 억제효과를 나타내었다. 그러나, 6일 동안 더 장기간의 프로게스테론 처리는 4일 동안의 처리기간에서 관찰된 세포증식에 대한 농도-의존성 억제효과를 제거하였다. 그리고, 타목시펜은 HeLa 세포주의 증식에 대한 억제효과를 위하여 에스트로겐 보다 더 높은 농도($100{\mu}g$/ml)를 필요로 하였다. 이러한 결과는 고농도의 에스트로겐, 프로게스테론 그리고 타목시펜이 HeLa 세포의 증식을 억제할 뿐만 아니라, 농도 및 처리기간 또한 세포증식에 대한 억제효과에 영향을 미칠 수 있음을 제시한다.

Keywords

References

  1. Adami, H. O., C. C. Hsieh, M. Lambe, D. Trichopoulos, D. Leon, I. Persson, A. Ekbom, and P. O. Janson. 1994. Parity, age at first childbirth, and risk of ovarian cancer. Lancet 344, 1250-1254. https://doi.org/10.1016/S0140-6736(94)90749-8
  2. Arbeit, J. M., P. M. Howley, and D. Hanahan. 1996. Chronic estrogen-induced cervical and vaginal squamous carcinogenesis in human papillomavirus type 16 transgenic mice. Proc. Natl. Acad. Sci. 93, 2930-2935. https://doi.org/10.1073/pnas.93.7.2930
  3. Bardin, A., N. Boulle, G. Lazennec, F. Vignon, and P. Pojol. 2004. Loss of $ER\beta$ expression as a common step in estrogen -dependent tumor progression. Endocrine Related Cancer 11, 537-551. https://doi.org/10.1677/erc.1.00800
  4. Brake, T., and P. F. Lambert. 2005. Estrogen contributes to the onset, persistence, and malignant progression of cervical cancer in a human papillomavirus-transgenic mouse model. Proc. Natl. Acad. Sci. 102, 2490-2495. https://doi.org/10.1073/pnas.0409883102
  5. Bu, S. Z., D. L.Yin, X. H. Ren, L. Z. Jiang, Z. J. Wu, Q. R. Gao, and G. Pei. 1997. Progesterone induces apoptosis and up-regulation of p53 expression in human ovarian carcinoma cell lines. Cancer 79, 1944-1950. https://doi.org/10.1002/(SICI)1097-0142(19970515)79:10<1944::AID-CNCR15>3.0.CO;2-V
  6. Chung, S. H., S. Franceschi, and P. F. Lambert1. 2010. Estrogen and $ER\alpha$: Culprits in Cervical Cancer? Trends Endocrinol. Metab. 21, 504-511. https://doi.org/10.1016/j.tem.2010.03.005
  7. Deroo, B. J. and K. S. Korach. 2006. Estrogen receptors and human disease. J. Clin. Invest. 116, 561-570. https://doi.org/10.1172/JCI27987
  8. EL-Ashry, D., D. A. Miller, S. Kharbanda, M. E. Lippman, and F. G. Kern. 1997. Constitutive Raf-1 kinase activity in breast cancer cells induces both estrogen-independent growth and apoptosis. Oncogene 15, 423-435. https://doi.org/10.1038/sj.onc.1201198
  9. Horvath, L. G., S. M. Henshall, C. S. Lee, D. R. Head, D. I. Quinn, S. Makela, W. Delprado, D. Golovsky, P. C. Brenner, G. O'Neill, R. Kooner, P. D. Stricker, J. J. Grygiel, J. A. Gustaffson, and R. L. Sutherland. 2001. Frequent loss of estrogen receptor-$\beta$ receptor expression in prostate cancer. Cancer Res. 61, 5331-5335.
  10. Hu, Z. and X. Deng. 2000. The effect of progesterone on proliferation and apoptosis in ovarian cancer cell. Zhonghua Fu Chan Ke Za Zhi. 35, 423-426.
  11. Imamov, O., G. J. Shim, M. Waner, and J. A. Gustafsson. 2005. Estrogen receptor beta in health and disease. Biol. Reprod. 73, 866-871. https://doi.org/10.1095/biolreprod.105.043497
  12. Koehler, K. F., L. A. Helguero, L. A. Haldosen, M. Warner, and J. A. Gustafsson. 2005. Refelections on the discovery and significance of estrogen receptor $\beta$. Endocrine Reviews 26, 465-478. https://doi.org/10.1210/er.2004-0027
  13. Konstantinopoulos, P. A., A. Kominea, G. Vandoros, G. P. Sykiotis, P. Andricopoulos, I. Varakis, G. Sotiropoulos- Bonikou, and A. G. Papavassiliou. 2003. Oestrogen receptor beta ($ER\beta$) is abundantly expressed in normal colonic mucosa, but declines in colon adenocarcinoma paralleling the tumor's dedifferentiation. European J. Cancer 39, 1251-1258. https://doi.org/10.1016/S0959-8049(03)00239-9
  14. Lambe, M., J. Wuu, M. A. Rossing, and C. C. Hsieh. 1999. Twinning and maternal risk of ovarian cancer. Lancet 353, 1941.
  15. Lewis-Wambi, J. S. and V. C. Jordan. 2009. Estrogen regulation of apoptosis: how can one hormone stimulate and inhibit? Breast Cancer Res. 11, 206. https://doi.org/10.1186/bcr2255
  16. Lonard, D. M. and Smith C. L. 2002. Molecular perspectives on selective estrogen receptor modulators (SERMs); progress in understanding their tissue-specific agonist and antagonist actions. Steroids 67, 15-24. https://doi.org/10.1016/S0039-128X(01)00133-7
  17. Nguyen, H. and V. Syed. 2011. Progesterone inhibits growth and induces apoptosis in cancer cells through modulation of reactive oxygen species. Gynecol. Endocrinol. 27, 830-836. https://doi.org/10.3109/09513590.2010.538100
  18. Nilsen, J., G. Mor, and F. Nafrolin. 2000. Estrogen-regulated developmental neuronal apoptosis is determined by estrogen receptor subtype and the Fas/Fas ligand system. J. Neurobiol. 43, 64-78. https://doi.org/10.1002/(SICI)1097-4695(200004)43:1<64::AID-NEU6>3.0.CO;2-7
  19. Petterson, K., K. Grandien, G. G. Kuiper, and J. A. Gustafsson. 1997. Mouse estrogen receptor $\beta$ forms estrogen response element-binding heterodimers with estrogen receptor $\alpha$. Mol. Endocrinol. 11, 1486-1496. https://doi.org/10.1210/me.11.10.1486
  20. Rho, Y. S. and G. Min. 2005. Relaxin does not influence the growth of human cervical adenocarcinoma (HeLa) cells in culture. Ann. N. Y. Acad. Sci. 1041, 470-480. https://doi.org/10.1196/annals.1282.071
  21. Riley, R. R, S. Duensing, T. Brake, K. Münger, P. F. Lambert, and J. M. Arbeit. Dissection of human papillomavirus E6 and E7 function in transgenic mouse models of cervical carcinogenesis. 2003. Cancer Res. 63, 4862-4871.
  22. Robertson, C. N., K. M. Roberson, G. M. Padilla, E. T. O'Brien, J. M. Cook, C. S. Kim, and R. L. Fine. 1996. Induction of apoptosis by diethylstilbestrol in hormone-insensitive prostate cancer cells. J. Natl. Cancer. Inst. 88, 908-917. https://doi.org/10.1093/jnci/88.13.908
  23. Rodriguez, G. C., N. P. Nagarsheth, K. L. Lee, R. C. Bentley, D. K. Walmer, M. Cline, R. S. Whitaker, P. Isner, A. Berchuck, R. K. Dodge, and C. L. Hughes. 2002. Progestin-induced apoptosis in the Macaque ovarian epithelium: differential regulation of transforming growth factor- beta. J. Natl. Cancer Inst. 94, 50-60. https://doi.org/10.1093/jnci/94.1.50
  24. Roger, P., M. E. Sahla, S. Makela, J. A. Gustafsson, P. Baldet, and H. Rochefort. 2001. Decreased expression of estrogen receptor $\beta$ protein in proliferative preinvasive mammary tumors. Cancer Res. 61, 2537-2541.
  25. Shai, A., A. T. Brake, C. Somoza, and P. F. Lambert. 2007. The human papillomavirus E6 oncogene dysregulates the cell cycle and contributes to cervical carcinogenesis through two independent activities. Cancer Res. 67, 1626-1635. https://doi.org/10.1158/0008-5472.CAN-06-3344
  26. Shim, W. S., M. Conaway, S. Masamura, W. Yue, J. P. Wang, R. Kmar, and R. J. Santen. 2000. Estradiol hypersensitivity and mitogen-activated protein kinase expression in long-term estrogen deprived human breast cancer cells in vivo. Endocrinol. 141, 396-405. https://doi.org/10.1210/en.141.1.396
  27. Song, R. X., G. Mor, F. Naftolin, R. A. McPherson, J. Song, Z. Zhang, W. Yue, J. Wang, and R. J. Santen. 2001. Effect of long-term estrogen deprivation on apoptotic responses of breast cancer cells to 17beta-estradiol. J. Natl. Cancer. Inst. 93, 1714-1723. https://doi.org/10.1093/jnci/93.22.1714
  28. Song, R. X. D. and R. J. Santen. 2003. Apoptotic action of estrogen. Apoptosis 8, 55-60. https://doi.org/10.1023/A:1021649019025
  29. Syed, V., G. Ulinski, S. C. Mok, G. K. Yiu, and S. M. Ho. 2001. Expression of gonadotropin receptor and growth responses to key reproductive hormones in normal and malignant human ovarian surface epithelial cells. Cancer Res. 61, 6768-6776.
  30. Syed, V. and S. M. Ho. 2003. Progesterone-induced apoptosis in immortalized normal and malignant human ovarian surface epithelial cells involves enhanced expression of Fas1. Oncogene 22, 6993-6890.
  31. Wada-Hiraike, O., O. Imamove, H. Hiraike, K. Hultenby, T. Schwend, Y. Omoto, M. Warner, and J. A. Gustafsson. 2006. Role of estrogen receptor $\beta$ in colonic epithelium. Proc. Natl. Acad. Sci. 103, 2959-2964. https://doi.org/10.1073/pnas.0511271103
  32. Yao, K., E. S. Lee, D. J. Bentrem, G. England, J. I. Schafer, R. M. O'Regan, and V. C. Jordan. 2000. Antitumor action of physiological estradiol on tamoxifen-stimulated breast tumors grown in athymic mice. Clin. Cancer. Res. 6, 2028-2036.
  33. Zhang, C. C. and D. J. Shapiro. 2000. Activation of the p38 mitogen-activated protein kinase pathway by estrogen or by 4-hydroxytamoxifen is coupled to estrogen receptor-induced apoptosis. J. Biol. Chem. 275, 479-486. https://doi.org/10.1074/jbc.275.1.479

Cited by

  1. Effective Anti-aging Strategies in an Era of Super-aging vol.20, pp.3, 2014, https://doi.org/10.6118/jmm.2014.20.3.85
  2. Fulvestrant Does Not Have Antagonistic Effect on 17β-estradiol's Anti-proliferative Action in Cultured Chinese Hamster Ovarian Cell Line vol.24, pp.2, 2014, https://doi.org/10.5352/JLS.2014.24.2.173