转录因子NFIC在cAMP信号通路调控根尖牙乳头干细胞分化中的作用

Role of NFIC on cAMP-mediated diferentiation of stem cells from the apical papilla

目的通过慢病毒转染过表达NFIC基因,研究NFIC在cAMP信号通路促进根尖牙乳头干细胞(SCAPs)分化中的作用。方法采用酶消化法培养SCAPs,将SCAPs分别于普通矿化诱导液(对照组)、cAMP信号通路激动剂(Forskolin组)、转染空病毒载体协同Forskolin(Forskolin+LV-empty组)、转染过表达NFIC慢病毒协同Forskolin(Forskolin+ovNFIC组)的矿化诱导液中培养。矿化诱导10 d后茜素红染色检测各组矿化结节形成情况,QPCR法检测7 d Runt相关基因2(RUNX2)、碱性磷酸酶(ALP)、骨钙素(OCN)mRNA的表达。结果 Forskolin作用SCAPs后矿化结节形成增多,相关矿化基因表达增高,而Forskolin+ov-NFIC组矿化结节进一步增多,矿化基因表达进一步上调。结论转录因子NFIC能够促进cAMP信号通路介导的SCAPs分化。

Objective To investigate the role of NFIC on the stimulation effects of cAMP-induced differentiation of stem cells from the apical papilla （SCAPs） in vitro. Methods SCAPs isolated from dental papilla of human imma- ture third molars were cultured by enzyme digestion. SCAPs were transfected with lentivirus that overexpressed NF- IC gene （ov-NFIC） or an empty vector （LV-empty） and co-treatment with Forskolin. Mineralized nodule formation of each group was measured by alizarin red staining. Quantitative real-time reverse-transcription polymerase chain reaction was performed to test the expressions of RUNχ2, ALP, OCN mRNA. Results Forskolin increased the ex- pression of Runχ2, ALP, OCN mRNA as well as matrix mineralization in SCAPs, and the stimulation effects of For- skolin were enhanced by overexpressing NFIC gene. Conclusion The results indicate that NFIC can promote cAMP-induced differentiation of SCAPs.