Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Silencing of Disabled-2 Gene by CpG Methylation in Human Breast Cancer Cell Line, MDA MB-231 Cells

사람의 유방암 세포주인 MDA MB-231 세포에서 CpG 메칠화에 의한 Disabled-2유전자의 발현억제

  • Ko Myung Hyun (Division of Biological Science, Wonkwang University) ;
  • Oh Yu Mi (Division of Biological Science, Wonkwang University) ;
  • Park Jun Ho (Division of Biological Science, Wonkwang University) ;
  • Jeon Byung Hoon (College of Oriental Medicine, Wonkwang University) ;
  • Han Dong Min (Division of Biological Science, Wonkwang University) ;
  • Kim Won Sin (Division of Biological Science, Wonkwang University)
  • 고명현 (원광대학교 자연대학 생명과학부) ;
  • 오유미 (원광대학교 자연대학 생명과학부) ;
  • 박준호 (원광대학교 자연대학 생명과학부) ;
  • 전병훈 (원광대학교 한의과대학 병리학교실) ;
  • 한동민 (원광대학교 자연대학 생명과학부) ;
  • 김원신 (원광대학교 자연대학 생명과학부)
  • Published : 2005.10.01
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Abstract

Human Disabled-2 (Dab2) is a candidate tumor suppressor gone that regulates cell growth by c-Fos suppression in normal cells. In many cancer cells, Dab2 expression is lost or greatly diminished in $\∼85\%$ of the breast and ovarian cancers. In this study, we have examined the methylation status of CpG island on Dab2 gene promoter using bisulfite-assisted genomic sequencing and methylation specific PCR (MSP) method in human breast cancer cell line, MDA MB-231 cells. In normal human uterus endometrial cells, Dab2 was completely unmethylated. In contrast, Dab2 was methylated on CpG dinucleotides near the TATA_ box in MDA MB-231 cells. following MDA MB-231 cells by treatment with 5-azacytidine, Dab2 gene were demethylated and reexpressed. Result of this study suggested that silencing of Dab2 gene is correlated to CpG island methylation in human breast cancer cell line, MBA MD-231 cells.

사람의 Disabled-2 (Dab2)는 정상세포에서 c-Fos의 발현을 억제하여 세포 성장을 조절하는 암억제 유전자로 추정되어 지고 있다. 많은 암세포에서 Dab2는 유방과 난소의 정상세포에서는 발현이 되지만, 약 $85\%$의 유방과 난소의 종양세포에서는 발현이 줄어들거나, 발현이 억제되는 것으로 알려져 있다. 본 연구에서는 bisulfite 반응에 의한 염기서열 분석법과 MSP방법 등을 이용하여 유방암 세포주인 MDA MB-231세포에서 Dab2 유전자의 promoter상에 존재하는 Cpg island의 methylation된 상태를 분석하였다. 그 결과, 사람의 정상 자궁내막세포에서는 Dab2 promoter 부위가 완전하게 methylation되어 있지 않았다. 그러나 MDA MB-231세포에서는 TATA box 근처 의 CpG dinucleotide에서 비정상적으로 methylation되어 있었다. 이런 비정상적인 CpG dinucleotide의 methylation은 MDA MB-231세포를 5-azacytidine으로 처리하였을 때 methylation이 풀리고, Dab2의 발현이 회복되는 것으로 나타났다. 따라서 인간 유방암 세포주인 MBA MB-231세포에서 Dab2의 발현억제는 Dab2 유전자의 promoter부위의 CpG island의 비정상적인 methylation과 관련이 있는 것으로 여겨진다.

Keywords

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