MicroRNA-135b inhibits odontoblast-like differentiation of human dental pulp cells by regulating Smad5 and Smad4
MicroRNA-135b inhibits odontoblast-like differentiation of human dental pulp cells by regulating Smad5 and Smad4
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MicroRNA-135b 通过调节 Smad5 和 Smad4 抑制人牙髓细胞的成牙本质细胞样分化。
DOI：
10.1111/iej.12678
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发表时间：
2017-07-01
影响因子：
5
通讯作者：
Tian, Y. G.
中科院分区：
文献类型：
--
作者：
Song, Z.;Chen, L. L.;Tian, Y. G.
目的探讨mirna在人牙髓细胞（hDPCs）成牙细胞样分化中的作用。方法采用综合比较miRNA微阵列分析来确定hDPCs在成牙细胞样分化过程中miRNA的差异表达。采用实时定量逆转录聚合酶链式反应（qRT-PCR）和原位杂交（ISH）技术检测miR-135b （miR-135b）的丰度。利用生物信息学分析结合荧光素酶测定来鉴定与miR-135b相互作用的靶标。通过过表达miR-135b来研究hDPCs在成牙细胞样分化中的作用和机制。统计分析采用单因素方差分析（ANOVA）或学生t检验。结果在hDPCs成牙细胞样分化过程中鉴定到36个差异表达的microrna。在hDPCs分化过程中，MiR-135b表达明显下调（P < 0.05）。此外，miR-135b能够结合Smad5和Smad4的30-UTR并抑制这两个基因的表达（P < 0.05）。此外，过表达miR-135b抑制了hDPCs的成牙细胞样分化，并减弱了Smad5和Smad4的表达（P < 0.05）。这些观察结果表明miR-135b在介导hDPCs成牙细胞样分化中的潜在作用，抑制miR-135b可能是促进牙本质组织工程的一种有希望的治疗方法。
Aim To investigate the function of miRNAs in odontoblast-like differentiation of human dental pulp cells (hDPCs).Methodology Integrated comparative miRNA microarray profiling was used to determine the differential miRNAs expression in odontoblast-like differentiation of hDPCs. The abundance of microRNA-135b (miR-135b) was measured by quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) and in situ hybridization (ISH). Bioinformatic analyses combined with luciferase assays were utilized to identify the targets interacting with miR-135b. Overexpression of miR-135b was performed to investigate the role and mechanism in odontoblast-like differentiation of hDPCs. Statistical analysis was performed by one-way analysis of variance (ANOVA) or Student's t-test.Results Thirty-six differentially expressed microRNAs in odontoblast-like differentiation of hDPCs were identified. MiR-135b expression was significantly downregulated during hDPCs differentiation (P < 0.05). In addition, miR-135b was able to bind to the 30-UTR of the Smad5 and Smad4 and repressed these two genes expression (P < 0.05). Furthermore, overexpression of miR-135b suppressed odontoblast-like differentiation of hDPCs and attenuated the expression of Smad5 and Smad4 (P < 0.05).Conclusions These observations indicated a potential role of miR-135b in mediating odontoblast-like differentiation of hDPCs and inhibition of miR-135b might be a promising therapeutic way to facilitate dentine tissue engineering.