Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Analysis of Differentially Expressed Genes by Resveratrol Using Membrane Microarray

Membrane Microarray를 이용한 Resveratrol에 의해 차별적으로 발현되는 유전자 군의 분석

  • Kim, Jong-Sik (Dept. of Biological Sciences, Andong National University) ;
  • Jang, Min-Jung (Dept. of Biological Sciences, Andong National University) ;
  • Kim, Hyo-Eun (Dept. of Biological Sciences, Andong National University) ;
  • Kim, Soon-Young (Dept. of Biological Sciences, Andong National University) ;
  • Kim, Byung-Oh (Dept. of Applied Biology, Sangju National University) ;
  • Sohn, Ho-Yong (Dept. of Food & Nutrition, Andong National University)
  • 김종식 (안동대학교 자연과학대학) ;
  • 장민정 (안동대학교 자연과학대학) ;
  • 김효은 (안동대학교 자연과학대학) ;
  • 김순영 (안동대학교 자연과학대학) ;
  • 김병오 (상주대학교 생명자원과학대학 생물응용학과) ;
  • 손호용 (안동대학교 생활과학대학 식품영양학과)
  • Published : 2007.08.30
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Abstract

In the present study, we investigated whether several phytochemicals (resveratrol, genistein, epicatechin gallate, dially disulfide, caffeic acid phenetyl ester) and sulindac sulfide could induce expression of tumor suppressor p53 protein in human colorectal HCT116 cells. We found that p53 was dramatically induced by all phytochemical treatments except sulindac sulfide. Among treated phytochemicals, we selected resveratrol for further experiments because it is one of the highest p53 inducer. Using a Western blot analysis, we found that resveratrol induced p53 in a dose- and time-dependent manner. Additionally, using membrane-based microarray analysis, we found that twenty-five genes were up-regulated and two genes were down-regulated by resveratrol treatment. Among the up-regulated genes, we selected 4 genes and performed reverse-transcription-PCR to confirm microarray data. The results of RT-PCR were highly accorded with those of membrane microarray. In addition, we found that thrombospondin-1 (TSP-1) expression was not dependent on p53 presence, whereas mammary serine protease inhibitor (MASPIN) expression was dependent on p53 expressed by resveratrol treatment. The results of this study may help to promote our understandings of the molecular mechanisms of chemoprevention that are mediated by resveratrol in human colorectal cancer.

본 연구에서는 다섯 종류의 phytochemical (resveratrol, genistein, epicatechin gallate, diaIly disulfide, caffeic acid phenetyl ester)과 sulindac sulfide가 암 억제 단백질 p53을 유도할 수 있는지에 대해 연구하였다. 처리한 모든 phytochemical에 의해 p53 단백질의 발현이 강하게 유도된 반면, sulindac sulfide에 의해서는 p53 단백질이 유도되지 않았다. 처리한 phytochemical 중 포도껍질이나 와인에 많이 들어있는 resveratrol에 의해 p53 단백질이 농도의존적 혹은 처리시간 의존적으로 증가 발현되는 것을 확인하였다. 암 억제 단백질인 p53 하위 단계의 유전자들만 집적되어 있는 membrane microarray를 이용하여 실험을 수행한 결과, 25개의 유전자가 up-regulation 된 반면, 2개의 유전자가 down-regulation 되는 것을 확인하였다. Up-regulation 되는 유전자중 4개를 선택하여, RT-PCR을 수행한 결과 모두 membrane microarray 실험의 결과와 일치하였다. 게다가 p53 null인 HCT116 세포주를 이용한 RT-PCR을 통하여 TSP-1 유전자의 발현은 p53 의존적이지 않은 반면, MASPIN 유전자는 p53 의존적임을 확인하였다. 이러한 연구 결과는 resveratrol에 의한 화학적 암 예방법의 분자생물학적 기전을 이해하는데 도움을 줄 것으로 기대된다.

Keywords

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