흰쥐 상아질모세포 분화과정에서 OD314의 역할
Role of OD314 During Odontoblast Differentiation
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간추림 : 상아질모세포는 상아질을 형성하고 유지하는 세포이다. 상아질모세포-특이 유전자로 널리 알려진 DSPP 는 상아질의 석회화 과정에는 중요한 역할을 하나, 현재까지 DSPP 이외의 인자를 동정하고 이를 통하여 상아질모 세포의 분화와 상아질의 석회화 과정을 분자생물학적으로 연구한 결과들은 거의 알려져 있지 않다. 본 연구에서는 최근에 상아질의 석회화 과정에 중요한 역할을 하는 것으로 알려진 OD314 유전자의 발현을 분석하고, OD314 유전자의 과발현과 발현억제가 상아질모세포의 분화에 미치는 영향을 연구하여 다음과 같은 결과를 얻었다.MDPC-23 세포의 분화과정에서 OD314 mRNA는 배양 시작부터 발현되기 시작하여 배양 21일까지 그 발현이 증가하였고, 배양 28일에도 강한 발현이 유지되었다. 면역세포형광 염색에서 OD314 단백질은 세포의 핵에서는 약하게 발현되었으나, 세포질에서 강하게 발현되었으며 특히 핵에 인접한 세포질에서 더욱 강한 발현을 보였다. 상아질모세포의 석회화 관련 유전자인 DSPP는 OD314의 발현을 억제하였을 때 발현이 현저히 증대 되었으며, ON은 OD314를 과발현 시켰을 때 발현이 감소하였다. 이상의 결과를 종합하면 OD314는 상아질모세포의 분화와 상아질의 형성 및 석회화 과정에 중요한 역할을 하는 새로운 인자로 사료된다.
Abstract
Odontoblasts are responsible for the formation and maintenance of dentin which is a mineralized part in dentin-pulp complex of tooth. OD314 was obtained by subtractive hybridization between odontoblasts and osteoblast/dental papilla cells, and differentiatially expressed in the odontoblasts but not in osteoblasts and dental papilla cells. In this study, to better understand the biological function of new odontoblast-enriched gene, OD314, we examined expression of OD314 in cultured MDPC-23 cells and intracellular localization of OD314 protein. We also evaluate the effect of OD314 over-expression and inactivation on the cells by northern analysis. When MDPC-23 cells are cultured in the differentiation and mineralization medium for 28 days, OD314 mRNA expression was gradually increased from the beginning to day 21 and remained relatively high on day 28. Immunofluorescent staining of cultured MDPC-23 revealed localization of OD314 on the cytoplasm, especially near the nuclear membrane. However, a small amount of fluorescence was also observed in the nucleus. Inactivation of OD314 by RNA interference up-regulated the expression of DSPP, whereas over-expression of OD314 by CMV-OD314 plasmid down-regulated the expression of ON. These results suggest that OD314, a odontoblat-enriched gene, may play important roles in the odontoblast differentiation and dentin mineralization.
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