Korean J Phys Anthropol.  2004 Jun;17(2):121-129.

Inactivation of the OD314 Gene by RNA Interference in Preodontoblast Cell Lines

• Kim HJ
• Jeong MJ
• Son HH
• Park JC

Abstract

Tooth development depends on reciprocal interactions between oral epithelium and ectomesenchyme. The ectomesenchyme-derived odontoblasts secrete several collagenous and non-collagenous proteins to form a unique extracellular matrix. OD314 was obtained by subtractive hybridization between odontoblasts and osteoblast/pulp cells, and differentially or predominantly expressed in odontoblasts of rat incisors compared to osteoblasts and pulp cells. However, little is known about the function of OD314 in odontoblast differentiation. In this study, to better understand the function of OD314, we inactivated the OD314 gene in mouse MDPC23 cells using U6 promoter-driven siRNA method. After the characterization of mineralized nodule formation and molecular expression of MDPC23 cell, Inactivation effects of the OD314 were evaluated by RT-PCR and western blot. 1. Mineralized nodule formation was observed after 14 days of culture in MDPC23 cells. 2. The OD314, OC and DSPP mRNA were highly expressed throughout the cultures, while the expression of ON decreased as the MDPC23 cells differentiated. 3. The OD314 protein was weakly expressed at 7 days of culture, but increased gradually as MDPC23 cells reached mineralization stage. 4. The inactivation of OD314 by RNA interference downregulated the expressions of OD314, DSPP, OC, and ON mRNA and OD314 protein in MDPC23 cells. These results suggest that OD314 may play a important role in mineralization process of odontoblast and also regulate odontoblast-related genes such as OC, ON, and DSPP.

Keyword

OD314; Odontoblast; SiRNA; Mineralization; Regulation

MeSH Terms

Animals
Blotting, Western
Cell Line*
Collagen
Epithelium
Extracellular Matrix
Incisor
Mice
Odontoblasts
Osteoblasts
Rats
RNA Interference*
RNA*
RNA, Messenger
RNA, Small Interfering
Tooth
Collagen
RNA
RNA, Messenger
RNA, Small Interfering