DOI QR코드

DOI QR Code

브로콜리 유래 sulforaphane에 의한 NAG-1과 p21 유전자의 발현 조절

Up-Regulation of NAG-1 and p21 Genes by Sulforaphane

  • 정병걸 ((주) 넨시스) ;
  • 김순영 (국립안동대학교 자연과학대학 생명과학과) ;
  • 이건주 (국립안동대학교 자연과학대학 생명과학과) ;
  • 김종식 (국립안동대학교 자연과학대학 생명과학과)
  • 투고 : 2011.11.17
  • 심사 : 2012.02.17
  • 발행 : 2012.03.30

초록

본 연구에서는 대장암 세포주 모델에서 브로콜리 유래 sulforaphane에 의한 항 성장 활성과 항암 단백질 NAG-1과 p21의 발현 및 발현 조절에 대해서 연구하였다. 그 결과, 처리한 sulforaphane의 농도가 증가됨에 따라 세포사멸이 증가되었고, 세포 생존율은 감소하였다. 또한, sulforaphane에 의한 항암 단백질 NAG-1과 p21의 발현증가를 농도별, 시간대별로 확인한 결과, 두 단백질 모두 sulforaphane 농도와 처리시간 의존적으로 발현이 증가됨을 확인하였다. Sulforaphane에 의한 NAG-1과 p21의 발현의 p53 의존성을 연구한 결과, p21의 발현은 p53에 의존적 이지만 NAG-1의 발현은 p53에 비 의존적인 것으로 생각된다. 또한, sulforaphane이 dietary histone deacetylase inhibitor로서 NAG-1의 발현을 증가시킬 가능성은 매우 미미한 것으로 생각된다. 전사조절인자인 ATF3의 발현을 sulforaphane을 시간대 별로 처리한 후 확인한 결과, 2시간째부터 발현이 증가되어 NAG-1의 발현 보다 먼저 증가됨을 확인하였다. 이러한 연구 결과는 sulforaphane의 항암 기능을 이해하는 새로운 기전을 제시해 줄 것으로 기대된다.

We investigated the anti-proliferative activity of sulforaphane and expression changes of NAG-1 and p21 genes in response to sulforaphane treatment in human colorectal HCT116 cells. The results showed that sulforaphane decreased cell viabilities in a dose-dependent manner and induced expression of NAG-1 and p21 proteins in a dose-dependent and time-dependent manner. In addition, we found that NAG-1 expression by sulforaphane was not dependent on the presence of p53, whereas p21 expression was dependent on p53 presence. The results indicated that up-regulation of NAG-1 was not related with the activity of a dietary histone deacetylase inhibitor of sulforaphane. ATF3 induction was detected from 2 hr after sulforaphane treatment, indicating that ATF3 could be a transcription factor to up-regulate NAG-1 expression. The results of this study may help to increase our understanding of the molecular mechanism of anti-cancer activity mediated by sulforaphane in human colorectal cancer cells.

키워드

참고문헌

  1. Baek, S. J., K. S. Kim, J. B. Nixon, L. C. Wilson, and T. E. Eling. 2001. Cyclooxygenase inhibitors regulate the expression of a TGF-beta superfamily member that has proapoptotic and antitumorigenic activities. Mol. Pharmacol. 59, 901-908.
  2. Baek, S. J., L. C. Wilson, and T. E. Eling. 2002. Resveratrol enhances the expression of non-steroidal anti-inflammatory drug-activated gene (NAG-1) by increasing the expression of p53. Carcinogenesis 23, 425-434. https://doi.org/10.1093/carcin/23.3.425
  3. Baek, S. J., J. S. Kim, F. R. Jackson, T. E. Eling, M. F. McEntee, and S. H. Lee. 2004. Epicatechin gallate-induced expression of NAG-1 is associated with growth inhibition and apoptosis in colon cancer cells. Carcinogenesis 25, 2425-2432. https://doi.org/10.1093/carcin/bgh255
  4. Baek, S. J., R. Okazaki, S. H. Lee, J. Martinez, J. S. Kim, K. Yamaguchi, Y. Mishina, D. W. Martin, A. Shoieb, M. F. McEntee, and T. E. Eling. 2006. Nonsteroidal anti-inflammatory drug-activated gene-1 over expression in transgenic mice suppresses intestinal neoplasia. Gastroenterology 131, 1553-1560. https://doi.org/10.1053/j.gastro.2006.09.015
  5. Berner, C., E. Aumüller, A. Gnauck, M. Nestelberger, A. Just, and A. G. Haslberger. 2010. Epigenetic control of estrogen receptor expression and tumor suppressor genes is modulated by bioactive food compounds. Ann. Nutr. Metab. 57, 183-189. https://doi.org/10.1159/000321514
  6. Brandenburg, L. O., M. Kipp, R. Lucius, T. Pufe, and C. J. Wruck. 2010. Sulforaphane suppresses LPS-induced inflammation in primary rat microglia. Inflamm. Res. 59, 443-450. https://doi.org/10.1007/s00011-009-0116-5
  7. Cekanova, M., S. H. Lee, R. L. Donnell, M. Sukhthankar, T. E. Eling, S. M. Fischer, and S. J. Baek. 2009. Nonsteroidal anti-inflammatory drug-activated gene-1 expression inhibits urethane-induced pulmonary tumorigenesis in transgenic mice. Cancer Prev. Res. 2, 450-458. https://doi.org/10.1158/1940-6207.CAPR-09-0057
  8. Dashwood, R. H. and E. Ho. 2007. Dietary histone deacetylase inhibitors: from cells to mice to man. Semin. Cancer Biol. 17, 363-369. https://doi.org/10.1016/j.semcancer.2007.04.001
  9. Fahey, J. W., Y. Zhang, and P. Talalay. 1997. Broccoli sprouts: an exceptionally rich source of inducers of enzymes that protect against chemical carcinogens. Proc. Natl. Acad. Sci. USA 94, 10367-10372. https://doi.org/10.1073/pnas.94.19.10367
  10. Ho, E., J. D. Clarke, and R. H. Dashwood. 2009. Dietary sulforaphane, a histone deacetylase inhibitor for cancer prevention. J. Nutr. 139, 2393-2396. https://doi.org/10.3945/jn.109.113332
  11. Jang, M. J., H. E. Kim, S. M. Son, M. J. Kim, E. W. Seo, Y. H. Kim, and J. S. Kim. 2009. Over-expression of NSAID activated gene-1 by caffeic acid phenethyl ester. J. Life Sci. 19, 1787-1793. https://doi.org/10.5352/JLS.2009.19.12.1787
  12. Kim, J. S., S. J. Baek, T. Sali, and T. E. Eling. 2005. The conventional nonsteroidal anti-inflammatory drug sulindac sulfide arrests ovarian cancer cell growth via the expression of NAG-1/MIC-1/GDF-15. Mol. Cancer Ther. 4, 487-493.
  13. Lee, S. H., J. S. Kim, K. Yamaguchi, T. E. Eling, and S. J. Baek. 2005. Indole-3-carbinol and 3,3'-diindolylmethane induce expression of NAG-1 in a p53-independent manner. Biochem. Biophys. Res. Commun. 328, 63-69. https://doi.org/10.1016/j.bbrc.2004.12.138
  14. Lee, S. H., K. Yamaguchi, J. S. Kim, T. E. Eling, S. Safe, Y. Park, and S. J. Baek. 2006. Conjugated linoleic acid stimulates an anti-tumorigenic protein NAG-1 in an isomer specific manner. Carcinogenesis 27, 972-981. https://doi.org/10.1093/carcin/bgi268
  15. Li, Y., T. Zhang, H. Korkaya, S. Liu, H. F. Lee, B. Newman, Y. Yu, S. G. Clouthier, S. J. Schwartz, M. S. Wicha, and D. Sun. 2010. Sulforaphane, a dietary component of broccoli/ broccoli sprouts, inhibits breast cancer stem cells. Clin. Cancer Res. 16, 2580-2590. https://doi.org/10.1158/1078-0432.CCR-09-2937
  16. Livak, K. J. and T. D. Schmittgen. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 25, 402-408. https://doi.org/10.1006/meth.2001.1262
  17. Meeran, S. M., A. Ahmed, and T. O. Tollefsbol. 2010. Epigenetic targets of bioactive dietary components for cancer prevention and therapy. Clin. Epigenetics 1, 101-116. https://doi.org/10.1007/s13148-010-0011-5
  18. Piyanuch, R., M. Sukhthankar, G. Wandee, and S. J. Baek. Berberine, a natural isoquinoline alkaloid, induces NAG-1 and ATF3 expression in human colorectal cancer cells. Cancer Lett. 258, 230-240. https://doi.org/10.1016/j.canlet.2007.09.007
  19. Sun, X., L. Mi, J. Liu, L. Song, F. L. Chung, and N. Gan. 2011. Sulforaphane prevents microcystin-LR-induced oxidative damage and apoptosis in BALB/c mice. Toxicol. Appl. Pharmacol. 255, 9-17. https://doi.org/10.1016/j.taap.2011.05.011
  20. Traka, M. H., K. F. Chambers, E. K. Lund, R. A. Goodlad, I. T. Johnson, and R. F. Mithen. 2009. Involvement of KLF4 in sulforaphane- and iberin-mediated induction of p21(waf1/cip1). Nutr. Cancer 61, 137-145. https://doi.org/10.1080/01635580802348641
  21. Wilson, L. C., S. J. Baek, A. Call, and T. E. Eling. Nonsteroidal anti-inflammatory drug-activated gene (NAG-1) is induced by genistein through the expression of p53 in colorectal cancer cells. Int. J. Cancer 105, 747-753. https://doi.org/10.1002/ijc.11173
  22. Yamaguchi, K., S. H. Lee, T. E. Eling, and S. J. Baek. 2006. A novel peroxisome proliferator-activated receptor gamma ligand, MCC-555, induces apoptosis via posttranscriptional regulation of NAG-1 in colorectal cancer cells. Mol. Cancer Ther. 5, 1352-1361. https://doi.org/10.1158/1535-7163.MCT-05-0528
  23. Yoshioka, H., H. Kamitani, T. Watanabe, and T. E. Eling. 2008. Nonsteroidal anti-inflammatory drug-activated gene (NAG-1/GDF15) expression is increased by the histone deacetylase inhibitor trichostatin A. J. Biol. Chem. 283, 33129-33137. https://doi.org/10.1074/jbc.M805248200

피인용 문헌

  1. TRAIL and Effect of Irradiation on Apoptosis of Cancer Cells vol.10, pp.6, 2016, https://doi.org/10.7742/jksr.2016.10.6.387