생명과학회지 (Journal of Life Science)

  • 제19권7호
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  • Pages.871-877
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  • 2009
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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Cdk inhibitors의 발현 증가 및 pRB 인산화 저해에 의한 HDAC inhibitor인 sodium butyrate에 의한 인체백혈병세포의 G1 arrest유발

G1 Arrest of U937 Human Monocytic Leukemia Cells by Sodium Butyrate, an HDAC Inhibitor, Via Induction of Cdk Inhibitors and Down-regulation of pRB Phosphorylation

  • 최영현 (동의대학교 한의과대학 생화학교실 및 대학원 바이오물질제어학과)
  • Choi, Yung-Hyun (Department of Biochemistry, Dongeui University College of Oriental Medicine and Department of Biomaterial Control, Dongeui University)
  • 발행 : 2009.07.30
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초록

대표적인 histone deacetylase inhibitor 저해제의 일종일 sodium butyrate에 의한 인체백혈병 U937세포의 증식 억제에 관한 기전 연구를 세포주기 조절 측면에서 조사하였다. MTT assay 및 flow cytometry 분석을 통하여 sodium butyrate의 처리 농도 증가에 따른 U937 세포의 증식억제는 세포주기 G1 arrest 및 apoptosis 유발에 의한 것임을 확인하였다. RT-PCR및 Western blotting 결과에서 sodium butrate에 의한 G1 arrest는 세포주기 G1기에서 S기로의 진입에 중요한 역할을 하는 cyclin D1, E, A, cyclin-dependent kinase (Cdk) 4 및 Cdk6발현의 저해와 p21 및 p27과 같은 Cdk inhibitor의 발현 증가와 연관성이 있었다. Sodium butyrate는 또한 retinoblastoma protein (pRB)및 p130 단백질의 인산화를 저해시켰으나, S기 진행에 중요한 전사조절인자인 E2F-1 및 E2F-4의 의 발현에는 큰 영향이 없었다. 그러나 sodium butyrate에 의한 pRB 및 p130단백질의 인산화 저해는 pRB와 E2F-1및 p130과 E2F-4와의 결합력을 증사시켰다. 본 연구의 결과는 U937세포의 증식억제에 pRB/p130 인산화 억제 및 Cdk inhibitors의 발현 증가가 중요한 역할을 하고있음을 보여주는 것으로, sodium butyrate의 항암기전 이해에 중요한 자료가 될 것이다.

We investigated the effects of sodium butyrate, a histone deacetylase inhibitor, on the cell cycle progression in human monocytic leukemia U937 cells. Exposure of U937 cells to sodium butyrate resulted in growth inhibition, G1 arrest of the cell cycle and induction of apoptosis in a dose-dependent manner as measured by MTT assay and flow cytometry analysis. The increase in G1 arrest was associated with the down-regulation in cyclin D1, E, A, cyclin-dependent kinase (Cdk) 4 and 6 expression, and up-regulation of Cdk inhibitors such as p21 and p27. Sodium butyrate treatment also inhibited the phosphorylation of retinoblastoma protein (pRB) and p130, however, the levels of transcription factors E2F-1 and E2F-4 were not markedly modulated. Furthermore, the down-regulation of phosphorylation of pRB and p130 by this compound was associated with enhanced binding of pRB and E2F-1, as well as p130 and E2F-4, respectively. Overall, the present results demonstrate a combined mechanism involving the inhibition of pRBjp130 phosphorylation and induction of Cdk inhibitors as targets for sodium butyrate that may explain some of its anti-cancer effects in U937 cells.

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참고문헌

  1. Archer, S. Y., J. Johnson, H. J. Kim, Q. Ma, H. Mou, V. Daesety, S. Meng, and R. A. Hodin. 2005. The histone deacetylase inhibitor butyrate downregulates cyclin B1 gene expression via a p21/WAF-1-dependent mechanism in human colon cancer cells. Am. J. Physiol. Gastrointest. Liver Physiol. 289, G696-703
  2. Baradari, V., A. Huether, M. Höpfner, D. Schuppan, and H. Scherübl. 2006. Antiproliferative and proapoptotic effects of histone deacetylase inhibitors on gastrointestinal neuroendocrine tumor cells. Endocr. Relat. Cancer 13, 1237-1250 https://doi.org/10.1677/erc.1.01249
  3. Candido, E. P., R. Reeves, and J. R. Davie. 1978. Sodium butyrate inhibits histone deacetylation in cultured cells. Cell 14, 105-113 https://doi.org/10.1016/0092-8674(78)90305-7
  4. Chopin, V., R. A. Toillon, N. Jouy, and X. Le Bourhis. 2004. P21(WAF1/CIP1) is dispensable for G1 arrest, but indispensable for apoptosis induced by sodium butyrate in MCF-7 breast cancer cells. Oncogene 23, 21-29 https://doi.org/10.1038/sj.onc.1207020
  5. Cummings, J. H. 1981. Short chain fatty acids in the human colon. Gut 22, 763-779 https://doi.org/10.1136/gut.22.9.763
  6. Danova, M., M. Giordano, and G. A. Mazzini. 1990. Expression of p53 protein during the cell cycle measured by flow cytometry in human leukemia. Leuk. Res. 14, 417-422 https://doi.org/10.1016/0145-2126(90)90027-7
  7. Elledges, S. J. and J. W. Harper. 1994. Cdk inhibitor: on the threshold of checkpoints and development. Curr. Opin. Cell Biol. 6, 847-852 https://doi.org/10.1016/0955-0674(94)90055-8
  8. Greenberg, V. L., J. M. Williams, J. P. Cogswell, M. Mendenhall, and S. G. Zimmer. 2001. Histone deacetylase inhibitors promote apoptosis and differential cell cycle arrest in anaplastic thyroid cancer cells. Thyroid 11, 315-325 https://doi.org/10.1089/10507250152039046
  9. Hara, I., H. Miyake, S. Hara, S. Arakawa, and S. Kamidono. 2000. Sodium butyrate induces apoptosis in human renal cell carcinoma cells and synergistically enhances their sensitivity to anti-Fas-mediated cytotoxicity. Int. J. Oncol. 17, 1213-1218
  10. Ibraghimov-Beskrovnaya, O. 2007. Targeting dysregulated cell cycle and apoptosis for polycystic kidney disease therapy. Cell Cycle 6, 776-779 https://doi.org/10.4161/cc.6.7.4047
  11. Johansson, M. and J. L. Persson. 2008. Cancer therapy: targeting cell cycle regulators. Anticancer Agents Med. Chem. 8, 723-731 https://doi.org/10.2174/187152008785914833
  12. Kruh, J. 1982. Effects of sodium butyrate, a new pharmacological agent, on cells in culture. Mol. Cell Biochem. 42, 65-82
  13. Li, X., M. Marani, R. Mannucci, B. Kinsey, F. Andriani, I. Nicoletti, L. Denner, and M. Marcelli. 2001. Overexpression of BCL-X (L) underlies the molecular basis for resistance to staurosporine-induced apoptosis in PC-3 cells. Cancer Res. 61, 1699-1706
  14. Litvak, D. A., K. O. Hwang, B. M. Evers, and C. M. Townsend. 2000. Induction of apoptosis in human gastric cancer by sodium butyrate. Anticancer Res. 20, 779-784
  15. Loo, D. T. and J. R. Rillema. 1998. Measurement of cell death. Methods Cell Biol. 57, 251-264 https://doi.org/10.1016/S0091-679X(08)61583-6
  16. Madigan, M. C., G. Chaudhri, P. L. Penfold, and R. M. Conway. 1999. Sodium butyrate modulates p53 and Bcl-2 expression in human retinoblastoma cell lines. Oncol. Rep. 7, 331-337
  17. Morgan, D. 1997. Cyclin dependent kinases: Engines, clocks and microprocessors. Annu. Rev. Cell Dev. Biol. 13, 261-291 https://doi.org/10.1146/annurev.cellbio.13.1.261
  18. Sandal, T. 2002. Molecular aspects of the mammalian cell cycle and cancer. Oncologist 7, 73-81 https://doi.org/10.1634/theoncologist.7-1-73
  19. Schwartz, B., C. Avivi-Green, and S. Polak-Charcon. 1998. Sodium butyrate induces retinoblastoma protein dephosphorylation, p16 expression and growth arrest of colon cancer cells. Mol. Cell Biochem. 188, 21-30 https://doi.org/10.1023/A:1006831330340
  20. Schwartz, G. K. and M. A. Shah. 2005. Targeting the cell cycle: a new approach to cancer therapy. J. Clin. Oncol. 23, 9408-9421 https://doi.org/10.1200/JCO.2005.01.5594
  21. Sherr, C. J. and J. M. Roberts. 1995. Inhibitors of mammalian G1 cyclin-dependent kinases. Genes Dev. 9, 1149-1163 https://doi.org/10.1101/gad.9.10.1149
  22. Sidle, A., C. Palaty, P. Dirks, O. Wiggan, M. Kiess, R. M. Gill, A. K. Wong, and P. A. Hamel. 1996. Activity of the retinoblastoma family proteins, pRB, p107, and p130, during cellular proliferation and differentiation. Crit. Rev. Biochem. Mol. Biol. 31, 237-271 https://doi.org/10.3109/10409239609106585
  23. Tsai, L. C., M. W. Hung, G. G. Chang, and T. C. Chang, 2000. Apoptosis induced by the sodium butyrate in human gastric cancer TMK-1 cells. Anticancer Res. 20, 2441-2448
  24. Vaziri, C., L. Stice, and D. V. Faller. 1998. Butyrate-induced G1 arrest results from p21-independent disruption of retinoblastoma protein-mediated signals. Cell Growth Differ. 9, 465-474
  25. Wang, A., R. Zeng, and H. Huang. 2008. Retinoic acid and sodium butyrate as cell cycle regulators in the treatment of oral squamous carcinoma cells. Oncol. Res. 17, 175-182 https://doi.org/10.3727/096504008785114129
  26. Wang, Y. F., N. S. Chen, Y. P. Chung, L. H. Chang, Y. H. Chiou, and C. Y. Chen. 2006. Sodium butyrate induces apoptosis and cell cycle arrest in primary effusion lymphoma cells independently of oxidative stress and p21(CIP1/WAF1) induction. Mol. Cell Biochem. 14, 1-9
  27. Weinberg, R. A. 1995. The retinoblastoma protein and cell cycle control. Cell 81, 323-330 https://doi.org/10.1016/0092-8674(95)90385-2
  28. Yen, A. and R. Sturgill. 1998. Hypophosphorylation of the RB protein in S and G2 as well as G1 during growth arrest. Exp. Cell Res. 241, 324-331 https://doi.org/10.1006/excr.1998.4007