探讨MSX1基因缺失导致的牙发育异常的机制及可能的信号通路  

To explore the mechanism and possible signal pathways of abnormal tooth development caused by MSX1 gene deletion

作　　者：周勇[1] 朱明会 黄旭瑶 邵海宾[1] 郑树灿[1] ZHOU Yong;ZHU Ming-hui;HUANG Xu-yao;SHAO Hai-bin;ZHENG Shu-can(Guangzhou Huadu District Maternal and Child Health Hospital,Affiliated Guangzhou Huadu Hospital,Guangdong Medical University,Guangdong Guangzhou 510800,China)

机构地区：[1]广州市花都区妇幼保健院广东医科大学附属广州花都医院,广东广州510000

出　　处：《临床口腔医学杂志》2021年第12期716-719,共4页Journal of Clinical Stomatology

摘　　要：目的:探讨MSX1基因缺失导致的牙发育异常的机制及可能的信号途径。方法:体外培养成牙本质细胞系MDPC-23,加入地塞米松(DEX),根据0、2、4、16、24 h时间节点完成细胞收集,采用荧光定量PCR测定细胞中MSX1、ALP、OC及MMP-2 mRNA表达水平。采用CRISP R-as9基因编辑技术对构建MSX1基因敲除小鼠成牙本质细胞进行精密编辑,设为观察组;选择成牙本质细胞系MDPC-23为对照组。采用实时荧光PCR和Wertern Blot法测定两组细胞中MSX1基因、ALP、OC及MMP-2蛋白水平。结果:成牙本质细胞系MC3T3-ET随着培养时间的延长MSX1基因、ALP、OC及MMP-2基因表达存在明显差异性。MSX1基因、ALP、OC及MMP-2基因培养12 h表达量均高于0、6、16及24 h(P<0.05)。观察组MSX1基因缺失后ALP、OC及MMP-2 mRNA水平均低于对照组(P<0.05);观察组ALP、OC及MMP-2蛋白水平明显下降,均低于对照组(P<0.05)。结论:MSX1基因缺失能引起牙发育异常,其机制可能与ALP、OC、MMP-2蛋白水平下调有关,从而引起相关信号通路途径异常。其机制可能与ALP、OC及MMP-2蛋白水平下调有关,从而引起相关信号通路途径异常。Objective:To investigate the mechanism and signal pathway of tooth dysplasia caused by Msx1 gene deletion.Methods:MDPC-23 cells were cultured in vitro and dexamethasone(DEX)was added.The cells were collected at 0,2,4,16 and 24 h time points.The contents of Msx1,ALP,OC and MMP-2 in the cells were determined by fluorescence quantitative PCR.The odontoblast cell line MDPC-23 of Msx1 knockout mice was constructed by CRISP R-Cas9 gene editing technology and set as the observation group,the odontoblast cell line MDPC-23 was selected as the control group.The expression of Msx1 gene,ALP,OC and MMP-2 Protein in the two groups were determined by real-time PCR and Wertern Blot.Results:The expression of Msx1,ALP,OC and MMP-2 in MC3 T3-ET was significantly different with the prolongation of culture time.The expression levels of Msx1,ALP,OC and MMP-2 in MC3 T3-ET cultured for 12 h were higher than those in 0,6,16 and 24 h(P<0.05).The mRNA levels of ALP,OC and MMP-2 in the observation group were lower than those in the control group(P<0.05).The protein levels of ALP,OC and MMP-2 in the observation group were significantly lower than those in the control group(P<0.05).Conclusion:MSX1 gene deletion can cause abnormal tooth development,and its mechanism may be related to the down-regulation of ALP,OC,and MMP-2 protein levels,resulting in abnormal signal pathways.The mechanism may be related to the down-regulation of ALP,OC and MMP-2 Protein Levels,resulting in abnormal signal pathways.

关 键 词：MSX1基因缺失 牙发育异常 信号途径 CRISP R-Cas9基因编辑技术 

分 类 号：Q813[生物学—生物工程]