DOI QR코드

DOI QR Code

Effects of dietary polyphenol (-)-epigallocatechin-3-gallate on the differentiation of mouse C2C12 myoblasts

식이성 폴리페놀 (-)-epigallocatechin-3-gallate가 mouse C2C12 myoblast 분화에 미치는 영향

  • Kim, Hye-Jin (Department of Exercise Science College of Health Science, Ewha Womans University) ;
  • Lee, Won-Jun (Department of Exercise Science College of Health Science, Ewha Womans University)
  • 김혜진 (이화여자대학교 건강과학대학 체육과학과) ;
  • 이원준 (이화여자대학교 건강과학대학 체육과학과)
  • Published : 2007.03.30

Abstract

In the present investigation, we studied the modulating effects of (-)-epigallocatechin-3-gallate(EGCG) on the differentiation of mouse C2C12 myoblasts. We found that the strong inhibitory effect of EGCG on DNA methyltransferase-mediated DNA methylation induced transdifferentiation of C2C12 myoblasts into smooth muscle cells demonstrated by both morphological changes and immunofluorescent staining. C2C12 myoblasts treated with EGCG for 4 days expressed smooth muscle ${\alpha}-actin$ protein. Real-time PCR data revealed that smooth muscle ${\alpha}-actin$ mRNA was induced by EGCG treated C2C12 myoblasts in a concentration-dependent manner. Smooth muscle ${\alpha}-actin$ mRNA concentration increased 330% and 490% after 2 and 3 days of 50 ${\mu}M$ of EGCG treatment. The expression of another smooth muscle marker, transgelin, mRNA was also increased up to 9-fold by 4 days of EGCG treatment compared with control in a concentration-dependent manner. These results suggested that C2C12 enables to transdifferentiate into smooth muscle when gene expression patterns are changed by the inhibition of DNA methylation induced by EGCG. In conclusion, transdifferentiation of C2C12 myoblasts into smooth muscle is resulted from the modulating effects of EGCG on DNA methylation which subsequently results in changing the expression pattern of several genes playing a critical role in the differentiation of C2C12 myoblasts.

본 연구에서는 유전자 발현에 중요한 조절 역할을 하는 DNA 메틸화를 식이성 폴리페놀의 하나인 녹차의 대표적인 추출물 EGCG로 억제하였을 때 C2C12 myoblast 세포에 일어나는 현상을 살펴보았다. 그 결과 smooth muscle의 지표인 transgelin, smooth muscle ${\alpha}-actin$ mRNA와 단백질이 현저히 증가함을 보였고, 형태학적으로도 smooth muscle의 전형적인 모습을 보였다. 식이에 포함된 DNA 메틸화 억제제인 EGCG가 C2C12 myoblast를 smooth muscle로 분화를 유도하였으며, 암 예방 차원에서의 EGCG의 역할 외에 혈관질환과 같은 smooth muscle에 관련된 예방과 치료차원에서 EGCG의 역할이 있을 것으로 사료된다. 본 연구는 C2C12 myoblast를 smooth muscle로 유도하는 결정적인 신호전달 역할을 하는 유전자에 대한 연구는 수행하지 못하였다. 따라서 EGCG에 의해 변화되는 유전자에 대한 기전연구가 필요하다고 하겠다.

Keywords

References

  1. Beard, C., E. Li and R. Jaenisch, 1995. Loss of methylaiton activates Xist in somatic but not embryonic cells. Genes & Development 9, 2325-2334 https://doi.org/10.1101/gad.9.19.2325
  2. Cheng, J. C., C. B. Matsen, F. A. Gonzales, W. Ye, S. Greer, V. E. Marquez, P. A. Jones and E. W. Selker. 2003. Inhibition of DNA methylation and reactivation of silenced genes by zebularine. Journal of the National Cancer Institute 95, 399-409 https://doi.org/10.1093/jnci/95.5.399
  3. Esteller, M. 2002. CpG island hypermethylation and tumor suppressor genes: a booming present, a brighter future. Oncogene 21, 5427-5440 https://doi.org/10.1038/sj.onc.1205600
  4. Fang, M. Z., Y. Wang, N. Ai, Z. Hou, Y. Sun, H. Lu, W. Welsh and C. S. Yang. 2003. Tea polyphenol (-)-epigallocatechin-3-gallate inhibits DNA methyltransfererase and reactivates methylation-silenced gene in cancer cell lines. Cancer Research 63, 7563-7570
  5. Fanzani, A., F. Colombo, R. Giuliani, A. Preti and S. Marchesini. 2006. Insulin-like growth factor 1 signaling regulates cytosolic sialidase Neu 2 expression during myoblast differentiation and hypertrophy. FEBS Journal 273, 3709-3721 https://doi.org/10.1111/j.1742-4658.2006.05380.x
  6. Herman, J. G. 1999. Hypermethylation of tumor suppressor genes in cancer. Seminars in Cancer Biology 9, 359-367 https://doi.org/10.1006/scbi.1999.0138
  7. Herman J. G. and S. B. Baylin. 2004. Gene silencing in cancer in association with promoter hypermethylation. New England Journal of Medicine 349, 2042-2054
  8. Jones, P. A. and P. W. Laird. 1999. Cancer epigenetics comes of age. Nature Genetics 21, 163-167 https://doi.org/10.1038/5947
  9. Jones, P. A. and S. B. Baylin. 2002. The fundamental role of epigenetic events in cancer. Nature Review of Genetics 3, 415-428
  10. Kondo, T. 2006. Epigenetic alchemy for cell fate conversion. Current Opinion in Genetics & Development 16, 502-507 https://doi.org/10.1016/j.gde.2006.07.001
  11. Langen R. C., A. M. Schols, M. C. Kelder, J. L. Van Der Velden, E. F. Wouters and Y. M. Janssen-Heininger. 2002. Tumor necrosis factor-alpha inhibits myogenesis through redox dependent and -independent pathways. American Journal of Physiology-Cell Physiology 283, C714-C721 https://doi.org/10.1152/ajpcell.00418.2001
  12. Lee, W. J. and B. T. Zhu. 2005. Mechanisms for the inhibition of DNA methyltransferases by tea catechins and bioflavonoids. Molecular Pharmacology 68, 1018-1030 https://doi.org/10.1124/mol.104.008367
  13. Lee, W. J. and B. T. Zhu. 2006. Inhibition of DNA methylation by caffeic acid and chlorogenic acid, two common catechol-containing coffee polyphenols. Carcinogenesis 27, 269-277 https://doi.org/10.1093/carcin/bgi206
  14. Li, E., C. Beard and R. Jaenisch. 1993. Role for DNA methylation in genomic imprinting. Nature 336, 362-365 https://doi.org/10.1038/336362a0
  15. Manabe, I. and G. K. Owens. 2001. Recruitment of serum response factor and hyperacetylation of histones at smooth muscle-specific regulatory regions during differentiation of a novel P19-derived in vitro smooth muscle differentiation system. Circulation Research 88, 1127-1134 https://doi.org/10.1161/hh1101.091339
  16. Momparler, R. L. 2003. Cancer epigenetics. Oncogene 22, 6479-6483 https://doi.org/10.1038/sj.onc.1206774
  17. Park, I. H., E. Erbay, P. Nuzzi and J. Chen. 2005. Skeletal myocyte hypertrophy requires mTOR kinase and S6K1. Experimental Cell Research 309, 211-219 https://doi.org/10.1016/j.yexcr.2005.05.017
  18. Paz, M. F., M. F. Fraga, S. Avila, M. Guo, M. Pollan, J. G. Herman and M. Esteller. 2003. A systemic profile of DNA methylation in human cancer cell lines. Cancer Research 63, 1114-1121
  19. Schmelz, K., M. Wagner, B. Dorken and I. Tamm. 2005. 5-Aza-2'-deoxycytidine induces p21WAF expression by demethylation of p73 leading to p53-independent apoptosis in myeloid leukemia. International Journal of Cancer 114, 683-695 https://doi.org/10.1002/ijc.20797
  20. Schmittwolf, C., N. Kirchhof, A. Jauch, M. Durr, F. Harder, M. Zenke and A. M. Muller. 2005. in vivo haematopoietic activity is induced in neurosphere cells by chromatin-modifying agents. EMBO Journal 24, 554-556 https://doi.org/10.1038/sj.emboj.7600546
  21. Spangenburg, E. E., D. K. Bowles and F. W. Booth. 2004. Insulin-like growth factor-induced transcriptional activity of the skeletal alpha-actin gene is regulated by signaling mechani는 linked to voltage-gated calcium channels during myoblast differentiation. Endocrinology 145, 2054-2063 https://doi.org/10.1210/en.2003-1476
  22. Spin, J. M., S. Nallamshetty, R. Tabibiazar, E. A. Ashley, J. Y. King, M. Chen, P. S. Tsao and T. Quertermous. 2004. Transcriptional profiling of in vitro smooth muscle cell differentiation identifies specific patterns of gene and pathway activation. Physiological Genomics 19, 292-302 https://doi.org/10.1152/physiolgenomics.00148.2004
  23. Springer, M. L., C. R. Ozawa and H. M. Blau. 2002. Transient production of alpha-smooth muscle actin by skeletal myoblasts during differentiation in culture and following intramuscular implantation. Cell Motility and the Cytoskeleton 51, 177-186 https://doi.org/10.1002/cm.10022
  24. Woodbury, D., K. Reynolds and I. B. Black. 2002. Adult bone marrow stromal stem cells express germline, ectodermal, endodermal, and mesodermal genes prior to neurogenesis. Journal of Neuroscience Research 69, 908-917 https://doi.org/10.1002/jnr.10365
  25. Wu, H. and Y. E. Sun. 2006. Epigenetic regulation of stem cell differentiation. Pediatric Research 59, 21R-25R https://doi.org/10.1203/01.pdr.0000203565.76028.2a

Cited by

  1. Protective Effect of Ferments of Hot-water Extract Mixture from Rhodiola sachalinensis and Red Ginseng on Oxidative Stress-induced C2C12 Myoblast vol.26, pp.3, 2013, https://doi.org/10.9799/ksfan.2013.26.3.485