人牙髓细胞分化过程中DNA甲基胞嘧啶双加氧酶TET家族的表达

发布时间：2018-09-05 18:32

【摘要】：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,qR T-PCR法检测矿化诱导第0,7,14 d的TET家族mR NA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(P0.05),TET3表达量在不同细胞代数之间差异无显著性意义(P0.05)。细胞经矿化诱导后,TET1,TET2和TET3的mR NA和蛋白表达量均升高(P0.05)。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。
[Abstract]:Background: TET family can oxidize methylcytosine to hydroxymethyl cytosine, participate in DNA demethylation, regulate cell proliferation and differentiation, but its expression pattern in dental pulp cells is still unclear. Aim: to investigate the expression pattern of TET family during odontoblast differentiation. Methods: human dental pulp cells were isolated and cultured by enzyme digestion. The expression of TET family and the expression of TET family in dental pulp cell subculture (P1-P7 passage) were detected by immunofluorescence assay. The level of mR NA in TET family was detected by QR T-PCR method at 714 d after mineralization induction in the third generation of dental pulp cells. The protein expression was detected by Western Blot method. Results and ConclusionThe cell cytoplasm and nucleus of human dental pulp cells were expressed in the family of: TET. In vitro passage of dental pulp cells, the expression of Tet T 1 protein in P1-P6 cells increased with cell passage. (P0.05) there was no significant difference between different cell generations in the expression of Tet T 1 protein in P2 and P3 generation cells (P0.05). The expression of mR NA and protein in Tet _ 1, T _ 2 and TET3 increased after mineralization (P0.05). These results suggest that TET family may be involved in the process of odontoblast differentiation.
【作者单位】：中山大学光华口腔医学院·附属口腔医院牙体牙髓病科;广东省口腔医学重点实验室;
【基金】：国家自然科学基金项目(81170953)~~
【分类号】：R329.2

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