Genome-based Gene Expression Analysis of EGCG-mediated Cell Transformation Suppression Effect in Mouse Cell line Balb/c 3T3 A31-1-1

마우스세포주 Balb/c 3T3 A31-1-1에서 Epigallocatechin gallate(EGCG)의 세포암화 억제효과에 대한 유전자발현 해석

  • Jung, Ki-Kyung (National Institute of Toxicological Research, Korea Food and Drug Administration) ;
  • Suh, Soo-Kyung (National Institute of Toxicological Research, Korea Food and Drug Administration) ;
  • Kim, Tae-Gyun (National Institute of Toxicological Research, Korea Food and Drug Administration) ;
  • Park, Moon-Suk (National Institute of Toxicological Research, Korea Food and Drug Administration) ;
  • Lee, Woo-Sun (National Institute of Toxicological Research, Korea Food and Drug Administration) ;
  • Park, Sue-Nie (National Institute of Toxicological Research, Korea Food and Drug Administration) ;
  • Kim, Seung-Hee (National Institute of Toxicological Research, Korea Food and Drug Administration) ;
  • Jung, Hai-Kwan (National Institute of Toxicological Research, Korea Food and Drug Administration)
  • 정기경 (식품의약품안전청 국립독성연구원) ;
  • 서수경 (식품의약품안전청 국립독성연구원) ;
  • 김태균 (식품의약품안전청 국립독성연구원) ;
  • 박문숙 (식품의약품안전청 국립독성연구원) ;
  • 이우선 (식품의약품안전청 국립독성연구원) ;
  • 박순희 (식품의약품안전청 국립독성연구원) ;
  • 김승희 (식품의약품안전청 국립독성연구원) ;
  • 정해관 (식품의약품안전청 국립독성연구원)
  • Published : 2006.12.31


Previous studies showed that epigallocatechin gallate(EGCG) have substantial effects of suppressing the N-methyl-N'-nitro-N-nitrosoguanidine(MNNG)-initiated cell transformation process on the bases of foci formation frequency and loss of anchorage dependency. In this study we tried to clarify the molecular mechanism of suppressing the cell transformation process. Mouse cell line balb/c 3T3 A31-1-1 was exposed 2 days to MNNG followed by 15 days 12-O-tetradecanoylphorbol-13-acetate(TPA) treatment for our transformation process. EGCG was added after the time point of 24 hours exposure to TPA and incubated for 19 days. 2029 genes were selected in our transformation process that showed fold change value of 1.5 or more in the microarray gene expression analysis covering the mouse full genome. These genes were found to be involved mainly in the cell cycle pathway, focal adhesion, adherens junction, TGE-$\beta$ signaling, apoptosis, lysine degradation, insulin signaling, ECM-receptor interaction. Among the genes, we focused on the 631 genes(FC>0.5) reciprocally affected by EGCG treatment. Our study suggest that EGCG down-regulate the gene expressions of up stream signaling factors such as nemo like kinase with MAPK activity and PI3-Kinase, Ras GTPase and down stream factors such as cyclin D1, D2, H, T2, cdk6.



  1. Ahmad, N., Cheng, P., and Mukhtar, H. (2000) Cell cycle dysregulation by green tea polyphenol epigallocatechin-3-gallate. Biochem. Biophys. Res. Commun., 275, 328-334
  2. Bozyczko-Coyne, D. (2001) CEP-1347/KT-7515, an inhibitor of SAPK/JNK pathway activation, promotes survival and blocks multiple events associated with Abeta-induced cortical neuron apoptosis. J. Neurochem., 77(3), 849-863
  3. Bredesen, D.E., Rao, R.V., and Mehlen, P. (2006) Cell death in the nervous system. Nature, 19;443(7113), 796-802
  4. Chen, Z.P., Schell, J.B., Ho, C-T., and Chen, K.Y. (1998) Green tea epigallocatechin gallate shows a pronounced growth inhibitory effect on cancerous cells but not on their normal counterparts. Cancer Lett., 129, 173-179
  5. Fujiki, H., Yoshizawa, S., Horiuchi, T., Suganuma, M., Yatsunami, J., and Nishiwaki-Matsushima, R. (1992) Anticarcinogenic effects of (−)-epigallocatechin gallate. Prev. Med., 4, 503-509
  6. Fujiki, H., Suganuma, M., Okabe, S., Komori, A., Sueoka, E., and Sueoka, N. (1998) Japanese green tea as a cancer preventive in humans. Nutr. Rev., 54, S67-70
  7. Gupta, S., Ahmad, N., Nieminen, A-L., and Mukhtar, H. (2000) Growth inhibition, cell-cycle dysregulation, and induction of apoptosis by green tea constituent (−)-epigallocatechin 3-gallate in androgen-sensitive and androgen-insensitive human prostate carcinoma cells. Toxicol. Appl. Paharmacol., 164, 91- 96
  8. Guputa, S., Hussain, T., and Mukhtar, H. (2003) Molecular pathway for (−)-epigalocatechin-3-gallate induced cell cycle arrest and apoptosis of human prostate carcinoma cell. Arch. biochem.biophys., 410, 177-183
  9. Hirose, M., Mizoguchi, Y., Yaono, M., Tanaka, H., Yamaguchi, T., and Shirai, T. (1997) Effects of green tea catechins on the progression or late promotion stage of mammary gland carcinogenesis in female sprague-Dawley rats pretreated with 7, 12 dimethylbenz (a) anthracene. Cancer Lett., 112, 141-147
  10. IARC (1985) IARC/NCI/EPA Working Group Cellular and molecular mechanisms of cell transformation and standardization of transformation assays of established cell lines for the prediction of carcinogenic chemicals: overview and recommended protocols. Cancer Research, 45, 2395-2399
  11. Kajiwara, Y., Ajimi, S., Hosokawa, A., Ajimi, S., Hosokawa, A., and Maekawa, K. (1997) Improvement of carcinogen detection in the BALB/3T3 cell transformation assay by using a rich basal medium supplemented with low concentration of serum and some growth factors. Mutation Research, 393, 81- 90
  12. Koh, S.H., Kwon, H., Kim, K.S., Kim, J., Kim, M.H., Yu, H.J., Kim, M., Lee, K.W., Do, B.R., Jung, H.K., Yang, K.W., and Appel, S.H., Kim, S.H. (2004) Epigallocatechin gallate prevents oxidative stress induced death of mutant Cu/Zn-superoxide dismutase (G93A) motoneuron cells by alteration of cell survival and death signals. Toxicology, 202(3), 213-225
  13. Lanfrancone, L., Pelicci, G., and Pelicci, P.G. (1994) Cancer genetics. Curr. Opin.. Genet. Dev., Feb; 4(1), 109-119
  14. LeBoeuf, R.A., Kerckaert, G.A., Aardema, M.J., Gibson, D.P., Brauninger, R., and Isfort, R.J. (1996) The pH 6.7 Syrian hamster embryo cell transformation assay for assessing the carcinogenic potential of chemicals. Mutation Research, 356, 85- 127
  15. Liao, S., Umekita, Y., Guo, J., Kokontis, J.M., and Hiipakka, R.A. (1995) Growth inhibition and regression of human prostate and breast tumors in athymic mice by tea epigallocatechin gallate. Cancer Lett., 96, 239-243
  16. Mukhtar, H. and Ahmad, N. (1999) Green tea in chemopreventon of cancer. Toxicol. Sci. [suppl], 52, 111-117
  17. Nasasni, I., Oh-hashi, F., Oh-hara, T., Feng, W.Y., Johnston, J., and Chan, K. (2003) Blocking telomerase by dietary polyphenols is a major mechanism for limiting the growth of human cancer cells in vitro and in vivo. Cancer Res., 63, 824-830
  18. Park, J.Y., Kwon, H.S., Yoon, C.Y., Park, M.S., Kim, B.H., Kim, J.S., Kang, H.I., and Jung, H.K. (2005) Screening of anticarcinogenic agents and their mode of action. The Annual Report of KFDA, 8(1), 238-245, 2004
  19. Senderowicz, A.M. (2004) Targeting cell cycle and apoptosis for the treatment of human malignancies. Curr. Opin. Cell Biol., 16(6), 670-678
  20. Tsuchiya, T. and Umeda, M., (1995) Improvement in the efficiency of the in vitro transformation assay method using BALB/3t3 A 31-1-1 cells. Carcinogenesis, 16, 1887-1894
  21. Vivanco, I. and Sawyers, C.L. (2002) The phosphatidylinositol 3- Klnase AKT pathway in human cancer., Nat Rev Cancer. 2(7), 489-501
  22. Yang, G-Y., Liao, J., Kim, K., Yurkow, E.J., and Yang, C.S. (1998) Inhibition of growth and induction of apoptosis in human cancer cell lines by tea polyphenol. Carcinogenesis, 19, 611-616
  23. Zhang, G.Y. (2005) Agents targeting c-Jun N-terminal kinase pathway as potential neuroprotectants. Expert Opin. Investig. Drugs, 14(11), 1373-1383