| 0811 Expression of Apin protein, OD314 , during ameloblast differentiation and mineralization | ||
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J.-T. PARK1, K.-Y. LEE1, H.-J. KIM1, H.H. SON2, S.-H. BAEK2, M.-I. CHO3, and J.-C. PARK1, 1Chosun university, College of Dentistry, Gwangju, South Korea, 2School of Dentistry, Seoul National University, Seoul National University Dental Hospital, South Korea, 3State University of New York - Buffalo, School of Dental Medicine, USA Objectives: Apin protein was isolated from CEOTs (calcifying epithelial odontogenic tumors)-associated amyloid and similar in sequence to OD314 by Dey et al (2001). This study aimed to uncover the biological function of Apin protein relating to ameloblast differentiation and mineralization in vitro. Methods: Ameloblast-lineage cells (Nakata et al, 2003) were prepared in minimum essential medium supplemented with 10% FBS, and 10ng/ml recombinant human EGF on type I collagen coated culture dishes. We examined the expression of Apin and several enamel matrix protein mRNAs during ameloblast differentiation and mineralization by RT-PCR. After inactivation and over-expression of Apin gene in ameloblast cell line the expression pattern of Apin and enamel matrix proteins were also evaluated by real time PCR and western analysis. Results: 1. When ameloblast cells were cultured in the differentiation and mineralization medium for 28 days, Apin protein expression was gradually increased from the beginning to day 28. 2. When ameloblast cells were cultured in the differentiation and mineralization medium for 28 days, the tuftelin mRNA expression was maintained from the beginning to day 14, and then gradually decreased to day 28. The expression of amelogenin and enamelin was gradually decreased according to the ameloblast differentiation. 3. Inactivation of Apin by U6-Apin SiRNA construct down-regulated the expression of Apin, MMP-20, and tuftelin, whereas over-expression of Apin by CMV-Apin construct up-regulated the expression of Apin and MMP-20 without change in tuftelin. Conclusion: The results suggest that Apin is considered as an amloblast-enriched protein and may play important roles in ameloblast differentiation and mineralization. This work was supported by RO1-2003-000-10141-0 from the Basic Research Program of The Korea Science & Engineering Foundation. | ||
| Seq #70 - Mechanisms of Odontogenesis 11:00 AM-12:00 PM, Thursday, 29 June 2006 Brisbane Convention & Exhibition Centre Exhibit Hall 1 | ||
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