In vitro expression of family with sequence similarity member 20C in human dental pulp cells and the effect of odontogenic induction on its expression

Abstract

Objective To investigate the expression pattern of family with sequence similarity member 20C (FAM20C) in human dental pulp cells (hDPCs) during subculture in vitro and the effect of odontogenic differentiation induction on its expression. Methods Western blotting was used to detect the FAM20C expression pattern of hDPCs from P0 to P7. Then the hDPCs were treated with odontogenic medium 7 and 14 days, and the mRNA and protein level of FAM20C were analyzed by RT-PCR and Western blotting, respectively. The expression of FAM20C protein during subculture was analyzed with t test. The mRNA and protein level of FAM20C during odontogenic differentiation were evaluated by One-Way ANOVA. Cellular localization and expression feature of FAM20C protein were determined by immunofluorescence in hDPCs during odontogenic differentiation. Results Western blot analysis showed that FAM20C protein level increased at passage 2-3 and decreased at passage 4-5 (tP2=-10.177, PP2= 0.001; tP3=-18.242, PP3<0.001; tP4=-19.143, PP4<0.001; tP5=-7.452, PP5= 0.002; tP6= 1.357, PP6= 0.246; tP7= 1.099, PP7=0.334) . FAM20C mRNA and protein level were up-regulated after the odontogenic induction of 7 days and 14 days, as observed by RT-PCR and Western blotting (FFAM20C mRNA= 86.252, PFAM20C mRNA, 7 d<0.001, PFAM20C mRNA, 14 d<0.001; FFAM20C protein= 85.569, PFAM20C protein, 7 d= 0.002, PFAM20C protein, 14 d<0.001) . FAM20C was mainly presented in cytoplasm after the odontogenic induction and the cytoplasmic protein level of FAM20C was elevated. Conclusions The expression of FAM20C was presented in hDPCs. The expression of FAM20C in hDPCs increased after induction of odontogenic differentiation. FAM20C might play an important role in the odontogenic differentiation of hDPCs. Key words: Dental pulp; Cells, cultured; In vitro; FAM20C; Odontogenic differentiation