背景：DNA甲基胞嘧啶双加氧酶TET家族可将甲基胞嘧啶氧化为羟甲基胞嘧啶，参与机体DNA去甲基化，调控细胞增殖与分化，但其在牙髓细胞中的表达模式仍不清楚。目的：检测TET家族在牙髓细胞成牙本质分化过程中的表达模式。方法：采用酶消化法分离培养人牙髓细胞，细胞免疫荧光法检测TET家族的表达及分布；Western Blot检测TET家族在牙髓细胞传代培养(P1-P7代)中的表达规律；对第3代牙髓细胞矿化诱导7，14 d，qRT-PCR法检测矿化诱导第0，7，14 d的TET家族mRNA水平，Western Blot法检测其蛋白量的表达变化。结果与结论：TET家族在人牙髓细胞的细胞质和细胞核中均有表达。在牙髓细胞体外传代培养过程中，TET1蛋白在P1-P6代随细胞传代呈上升趋势，TET2蛋白在P2和P3代细胞中表达增强(P<0.05)，TET3表达量在不同细胞代数之间差异无显著性意义(P >0.05)。细胞经矿化诱导后，TET1，TET2和TET3的mRNA和蛋白表达量均升高(P<0.05)。以上结果提示TET家族可能参与调控牙髓细胞成牙本质分化进程。%BACKGROUND:Ten-eleven translocation (TET) family proteins are recently discovered DNA dioxygenases that convert methylcytosine to hydroxymethyl cytosine, which is essential for regulating cel proliferation and differentiation, but the expression pattern of TET family proteins in human dental pulp cel s is stil unclear. OBJECTIVE:To investigate the expression pattern of TET family proteins during the differentiation of human dental pulp cel s. METHODS:Cel ular distribution and expression of TET family proteins were determined by immunofluorescence in human dental pulp cel s that were cultured and isolated using digestion method. The protein levels of TETs during cel passage (P1-P7) were detected with western blot assay, and their potential changes during odontogenic induction (7 and 14 days) were confirmed using real-time quantitative PCR and western blot analyses at mRNA and protein levels, respectively. RESULTS AND CONCLUSION:Al TETs were expressed in the nucleus and the cytoplasm of human dental pulp cel s During serial cel passage, TET1 protein expression was increased until the 6th passage, TET2 significantly increased at the 2nd and 3rd passages and then decreased (P<0.05), and TET3 showed no statistical y significant change (P>0.05). Both mRNA and protein expression levels of al TETs were elevated during odontogenic induction (P<0.05). These results indicated that TETs may contribute to cel differentiation of human dental pulp cel s.
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