Journal of Nutrition and Health

The Korean Nutrition Society (한국영양학회)
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Effect of EGCG on Expression of Neurogenin 3 via the MAP Kinase Signaling Pathway in AR42J Cells, a Rat Pancreatic Tumor Cell Line

녹차 카테킨, Epigallocathechin Gallate (EGCG)의 흰쥐췌장종양 선 세포 AR42J의 MAP Kinase 세포 신호전달 기전을 통한 Neurogenin 3 발현에 미치는 영향

  • Kim, Sung-Ok (Department of surgery and the Sealy Center for Cancer Cell biology, University of Texas Medical Branch) ;
  • Choe, Won-Kyung (Department of Food Nutrition, Gimcheon University)
  • 김성옥 (텍사스의과대학교 실리 암센터) ;
  • 최원경 (김천대학교 식품영양학과)
  • Received : 2011.03.16
  • Accepted : 2011.05.29
  • Published : 2011.06.30
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Abstract

Epigallocatechin gallate (EGCG), or epigallocatechin 3-gallate, is the ester of epigallocatechin and gallic acid, and is a type of catechin. EGCG may be therapeutic for many disorders including diabetics and some types of cancer. However it is unknown whether EGCG can induce transdifferentiation of pancreatic cells in pancreatitis. The aim of this study was to investigate the effects of EGCG on the expression of pancreatic regenerating related markers in pancreatic AR42J cells, a model of pancreatic progenitor cells. AR42J cells, differentiated with betacellulin and activin A, were cultured with/without EGCG in a time-dependent manner. Cell growth rate, levels of mRNA, and protein expression were examined with the MTT assay, quantitative PCR, and Western blots, respectively. The results showed that AR42J cell growth rates were inhibited by EGCG in a dose-dependent manner. mRNA and protein expression of amylase, insulin and neurogenin 3 (ngn 3) increased in AR42J cells treated with EGCG. Additionally, we demonstrated that the signal transduction pathway of mitogen-activated protein (MAP) kinase is active in EGCG-treated AR42J cells. ERK and JNK phosphorylation decreased in cells treated with EGCG but not p38 phosphorylation. Activation of the p38 MAP kinase pathway was confirmed by specific MAP kinase pathways inhibitors: U0126 for ERK, SP600126 for JNK, and SB203580 for p38. Activated p38 phosphorylation was inhibited by the specific p38 inhibitor SB203580 but p38 phosphorylation was inhibited with increased EGCG treatment. The ERK and JNK MAP kinase pathways were not affected by EGCG treatment. Although further studies are needed, these results suggest that EGCG affects the induction of pancreatic cell regeneration by increasing the ngn 3 protein and mRNA expression and activating the p38 MAP kinase pathway.

본 연구는 EGCG의 항 당뇨 활성기전으로 췌장종양 선세포 AR42J의 분화 및 내분비기능 개선에 미치는 영향과 그 세포 신호전달 기전을 확인하였다. 그 결과 첫째, AR42J 세포에 EGCG 처리 시 췌장종양 선세포의 세포증식이 농도 의존적으로 감소되었다. 둘째, 세포사멸 유도가 유의적으로 일어나지 않는 농도인 1uM EGCG를 AR42J 세포에 처리한 결과 ngn 3, ${\alpha}$-amylase, insulin은 EGCG처리 24시간에 mRNA, 단백질 발현증가를 나타내었고 48시간에 유의적 증가를 나타내었다. 셋째, EGCG 처리 시 ERK, JNK MAP Kinase 기전은 인산화 억제를 나타내었고 반면에 p38 기전의 인산화는 48시간에 유의적 증가를 하였다. 넷째, p38기전 저해제인 SB203580을 처리하여 EGCG가 MAP Kinase 기전중의 하나인 p38 기전 인산화 활성의 회복을 나타내어 ngn 3 발현을 위한 전사 신호전달 기전임을 다시 확인하였다. 따라서 녹차 생리활성 성분인 EGCG의 췌장종양 선 세포 AR42J 처리 결과 EGCG는 p38 MAP Kinase 기전 활성을 통해 췌장 선세포의 분화지표인 ngn 3 발현을 증가시키며 췌장내분비 기능 지표인 ${\alpha}$-amylase, insulin 발현증가를 나타내어 세포의 내분비기능 개선에도 영향을 미치는 것으로 사료된다.

Keywords

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