锌指E盒结合同源框2在人牙髓组织和细胞的表达

Expression of the zinc-finger E-box binding homeobox 2 in human dental pulp tissue and cells

目的:检测锌指E盒结合同源框2(ZEB2)在人牙髓组织和细胞中的表达,并初步探讨其意义。方法:制作牙髓组织石蜡切片,荧光原位杂交技术(FISH)检测ZEB2的表达;实时荧光定量聚合酶链反应(RT-qPCR)检测正常及TGF-β1刺激下人牙髓细胞(hDPCs)中ZEB2 mRNA表达水平;制作hDPCs细胞爬片,FISH检测ZEB2的表达。结果:FISH结果显示ZEB2在牙髓组织中主要表达于成牙本质细胞层,在牙髓细胞呈弱阳性表达。RT-qPCR结果显示TGF-β1的作用促进ZEB2的表达,且具有浓度和时间依赖性,5ng/ml TGF-β1刺激ZEB2表达达最大(P<0.05);5ng/ml TGF-β1刺激后,ZEB2 mRNA在24h内表达逐渐上调,24h达峰值(P<0.05)。FISH结果示ZEB2在正常hDPCs胞质和胞核均呈弱阳性表达,TGF-β1刺激24h后ZEB2表达增强,胞核表达明显。结论:ZEB2在牙髓组织中主要表达于成牙本质细胞层,正常hDPCs中弱阳性表达,而TGF-β1可促进hDPCs中ZEB2表达,提示ZEB2可能通过参与TGF-β1信号通路调控hDPCs的成牙本质向分化过程。

Objective：To investigate the expression of zinc-finger E-box binding homeobox 2 in human dental pulp tissue and cells. Methods： Using fluorescence in situ hybridization （FISH） to test and verify the expression of ZEB2 in nomal dental pulp tissue. The expression levels of ZEB2 in normal hDPCs and hDPCs stimulated by TGF-β1 were detected by real-time fluorescentquantitative PCR （RT-qPCR） and FISH. Results： FISH array of pulp tissue showed that ZEB2 was mainly expressed in odontoblasts. The expression of ZEB2 was weakly positive in the nucleus and cytoplasm of normal hDPCs. TGF-β1 can promote the expression of ZEB2. The expression of ZEB2 was up-regulated obviously when hDPCs were stimulated by 5ng/ml TGF-β1. RT-PCR showed that ZEB2 mRNA tending to increase from 0h to 24h. There was a down trend a over time （P〈0.05）. After stimulated by TGF- β1 for 24h, the expression of ZEB2 was strong positive, especially in nuclear. Conclusion：ZEB2 was mainly expressed in odontoblasts in normal dental pulp tissue. The expression level of ZEB2 was positively correlated with TGF-β1 which suggested that ZEB2 involved in TGF-β1 signaling pathway and may play an role of regulation in the process of hDPCs differentiated into odontoblast.