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Analysis of p53-Dependency of Differentially Expressed Genes by Capsaicin in Human Colorectal Cancer Cell

인간 대장암 세포주에서 capsaicin 처리에 의한 차별적인 유전자 발현의 p53 의존성 분석

  • Kim, Hyo-Eun (Department of Biological Sciences, Andong National University) ;
  • Jang, Min-Jeong (Department of Biological Sciences, Andong National University) ;
  • Lim, Seung-Hyun (Department of Biological Sciences, Andong National University) ;
  • Kim, Hyo-Rim (Department of Biological Sciences, Andong National University) ;
  • Kim, Soon-Young (Department of Biological Sciences, Andong National University) ;
  • Lee, Gun-Joo (Department of Biological Sciences, Andong National University) ;
  • Kim, Jong-Sik (Department of Biological Sciences, Andong National University)
  • 김효은 (안동대학교 자연과학대학 생명과학과) ;
  • 장민정 (안동대학교 자연과학대학 생명과학과) ;
  • 임승현 (안동대학교 자연과학대학 생명과학과) ;
  • 김효림 (안동대학교 자연과학대학 생명과학과) ;
  • 김순영 (안동대학교 자연과학대학 생명과학과) ;
  • 이건주 (안동대학교 자연과학대학 생명과학과) ;
  • 김종식 (안동대학교 자연과학대학 생명과학과)
  • Received : 2009.11.16
  • Accepted : 2009.11.30
  • Published : 2010.02.28

Abstract

In the present study, we investigated anti-proliferative activities of capsaicin and gene expression changes in response to capsaicin treatment in human colorectal HCT116 cells. The results showed that capsaicin decreased cell viabilities in a dose dependent manner and induced global gene expression changes. We found that 103 genes were up-regulated more than twofold, whereas 153 genes were down-regulated more than twofold by $100\;{\mu}M$ capsaicin treatment. Among the up-regulated genes, we selected 4 genes (NAG-1, DDIT3, GADD45A and PCK2) and performed RT-PCR to confirm the microarray data. We found that $100\;{\mu}M$ of capsaicin increased tumor suppressor p53 gene expression. In addition, the results showed that NAG-1, DDIT3 and GADD45A expressions were not dependent on p53 presence, whereas PCK2 expression. The results of this study may help to increase our understandings of the molecular mechanism of anti-proliferative activity mediated by capsaicin in human colorectal cancer cells.

본 연구에서는 대장암 세포주 모델에서 파이토케미칼 capsaicin에 의한 항 생장 활성과 유전체 수준에서의 유전자 발현 변화를 연구하였다. 그 결과, 처리한 capsaicin 농도 의존적으로 세포 생존율이 감소함을 확인하였고, capsaicin은 다양한 유전자의 발현 변화를 유도하였다. DNA microarray 실험결과 $100\;{\mu}M$ capsaicin의 처리에 의해 2배 이상 증가되는 유전자 103개가 확인된 반면, 2배 이상 발현이 감소되는 유전자 153개가 확인되었다. 발현이 증가되는 유전자 중 4개(NAG-1, DDIT3, GADD45A 그리고 PCK2)를 선택하여 RT-PCR을 수행한 결과, DNA micorarray 실험과 일치함을 확인하였다. 또한 $100\;{\mu}M$ capsaicin의 처리에 의해 암 억제유전자인 p53의 발현이 증가됨을 RT-PCR과 real-time PCR 방법으로 확인하였다. 게다가, NAG-1, DDIT3 그리고 GADD45A 유전자는 p53의 존재에 관계없이 발현이 증가되는 반면, PCK2 유전자는 반드시 p53에 의해 발현이 유도됨을 확인할 수 있었다. 이러한 연구는 대장암 세포주에서 capsaicin에 의한 항암 기전을 이해하는데 도움을 줄 것으로 기대된다.

Keywords

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