FAM20C在体外培养人牙髓细胞中的表达及成牙本质向分化诱导对其表达的影响

目的研究人牙髓细胞（hDPC）体外培养传代过程中FAM20C的表达模式，及对hDPC成牙本质向分化诱导后FAM20C的表达变化。 方法蛋白免疫印迹法（Western blot）检测第1代至第7代（P1 ~ P7）hDPC中FAM20C蛋白的表达量；结果采用独立样本t检验；对P3代hDPC进行成牙本质分化诱导7和14 d后，反转录聚合酶链反应与Western blot检测FAM20C的mRNA和蛋白水平变化。牙本质涎磷蛋白（DSPP）、牙本质基质蛋白1（DMP1）表达变化，FAM20C的mRNA以及蛋白表达变化采用单因素方差分析。免疫荧光法检测FAM20C在hDPC中的分布。 结果体外培养的hDPC中可检测到FAM20C表达，表达量随细胞传代先增加后减少（t_P2=-10.177，P_P2= 0.001；t_P3=-18.242，P_P3<0.001；t_P4=-19.143，P_P4<0.001；t_P5=-7.452，P_P5= 0.002；t_P6= 1.357，P_P6= 0.246；t_P7= 1.099，P_P7= 0.334）；成牙本质向分化诱导7和14 d后，FAM20C的mRNA和蛋白表达量高于未诱导组细胞（F_{FAM20C mRNA}=86.252，P_{FAM20C mRNA，7 d}<0.001，P_{FAM20C mRNA，14 d}<0.001；F_FAM20C蛋白= 85.569，P_{FAM20C蛋白，7 d}= 0.002，P_{FAM20C蛋白，14 d}<0.001）。免疫荧光结果显示，成牙本质诱导前FAM20C蛋白表达于细胞核，诱导后主要表达于细胞质，诱导14 d FAM20C在细胞质中表达量高于诱导7 d。 结论hDPC中表达FAM20C，成牙本质向分化诱导上调其表达水平，诱导后在细胞质中表达较高，提示FAM20C与hDPC的成牙本质分化相关。

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

ObjectiveTo 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. MethodsWestern 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. ResultsWestern blot analysis showed that FAM20C protein level increased at passage 2-3 and decreased at passage 4-5 (t_P2=-10.177, P_P2= 0.001; t_P3=-18.242, P_P3<0.001; t_P4=-19.143, P_P4<0.001; t_P5=-7.452, P_P5= 0.002; t_P6= 1.357, P_P6= 0.246; t_P7= 1.099, P_P7=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 (F_{FAM20C mRNA}= 86.252, P_{FAM20C mRNA, 7 d}<0.001, P_{FAM20C mRNA, 14 d}<0.001; F_{FAM20C protein}= 85.569, P_{FAM20C protein, 7 d}= 0.002, P_{FAM20C protein, 14 d}<0.001) . FAM20C was mainly presented in cytoplasm after the odontogenic induction and the cytoplasmic protein level of FAM20C was elevated. ConclusionsThe 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.