ODAM과 BMPRIB가 법랑질의 석회화에 미치는 영향

Effect of ODAM and BMPRIB on Enamel Mineralization

  • 박종태 (연세대학교 치과대학 구강생물학교실) ;
  • 조광희 (조선대학교 치과대학 구강해부학교실) ;
  • 배현숙 (남서울대학교 치위생학과) ;
  • 조영식 (남서울대학교 치위생학과) ;
  • 김흥중 (조선대학교 치과대학 구강해부학교실)
  • Park, Jong-Tae (Dept. of Oral Biology, Human Identification Research Center, Brain Korea 21 Project, Yonsei University College of Dentistry) ;
  • Cho, Kwang-Hee (Dept. of Oral Anatomy, School of Dentistry, Chosun University) ;
  • Bae, Hyun-Sook (Dept. of Dental Hygiene, Namseoul University) ;
  • Cho, Young-Sik (Dept. of Dental Hygiene, Namseoul University) ;
  • Kim, Heung-Joong (Dept. of Oral Anatomy, School of Dentistry, Chosun University)
  • 투고 : 2011.01.11
  • 심사 : 2011.02.24
  • 발행 : 2011.02.28

초록

최근에 Odontogenic ameloblast-associated protein (ODAM)은 MMP-20의 발현을 조절하여 법랑모세포 분화와 법랑질의 석회화에 중요한 역할을 한다고 보고되었다. 그러나 이에 대한 명확한 기전은 알려져 있지 않다. 그러므로 이 연구의 목적은 법랑모세포 분화와 법랑질의 석회화 과정에서의 ODAM의 생물학적 기능과 신호 전달 경로를 찾고자 하였다. Ameloblast-lineage cells (ALCs)를 이용하여 ODAM 재조합 단백질을 생성하고 ODAM 과발현 (ODAM overexpressing) 또는 ODAM 억제(ODAM silencing) 세포주를 만들었다. 세포들은 2주 동안 분화 배지에서 ODAM 재조합 단백질을 처리한 군과 처리하지 않는 군으로 나누어 배양하였다. ODAM의 신호 전달 경로를 확인하기 위하여, ALCs에 BMP2와 BMP receptor 1B (BMPR-1B) 억제제인 BAMBI 재조합 단백질을 처리하였고, 또한 BMPR-1B siRNA 이용하여 BMPR-1B의 발현을 억제하였다. 단백질 발현은 western blot 이용하여 분석하였다. 석회화는 sense ODAM 과발현 세포와 ODAM 재조합 단백질을 첨가한 법랑모세포 세포주에서 증진되었다. 또한 ALP 활성화는 sense ODAM 과발현 세포와 ODAM 정제된 단백질를 첨가한 법랑모세포 세포주에서 뚜렷하게 증진되었다. 기관발생과 관련이 있는 BMPR-IB와 석회화 과정과 관련된 CBP2는 ODAM 과발현을 유도한 경우에는 발현이 증가되었으나, ODAM 발현을 억제시킨 경우에는 발현이 현저히 감소하였다. 이상 실험의 결과는 법랑질 형성과정에서 ODAM이 법랑질 석회화를 증진시킬 수 있음을 시사한다.

The purpose of this study was to investigate the biological function of ODAM and its signal transduction pathway in the steps of ameloblast differentiation and enamel mineralization. An ODAM recombinant protein was produced and stable ODAM transgenic cell lines were also established using ameloblast-lineage cells (ALCs). To verify the ODAM signal transduction pathway, BAMBI recombinant protein, an inhibitor of BMP2 and BMP receptor 1B (BMPR-1B), was treated and BMPR-1B siRNA was used to silence expression of BMPR-1B. Mineralization was augmented by the ALCs treated with the ODAM recombinant protein and the sense ODAM overexpressing cells. The ALP activity was also increased markedly in the sense ODAM overexpressing cells and the ALCs treated with ODAM recombinant protein. The inactivation of ODAM in the ALCs down-regulated the expression of BMPR-1B, whereas its expression was up-regulated markedly when ODAM was overexpressed. These results provide deeper insights into the process of ameloblast maturation and in enamel mineralization. It also suggested that ODAM augmented enamel mineralization.

키워드

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