摘要
目的:研究EGF/EGFR/ErbB2信号通路对小鼠成釉细胞中基质金属蛋白酶-20(matrixmetalloproteinase-20,MMP-20)基因表达的调控作用。方法:用定时定量RT-PCR、双荧光素酶报告基因检测系统分析EGF/EGFR/ErbB2调控MMP-20基因的表达、EGF调控MMP-20启动子的主要区域、MAPK信号通路阻断剂对EGF调控MMP-20转录活性的影响、转录因子Jun家族介导EGF/ErbB2对MMP-20转录活性的调控;用基因定点突变和双荧光素酶基因检测报告系统分析EGF/ErbB2、C-Jun对MMP-20基因启动子转录活性的影响。结果:成釉细胞经20 ng/mL EGF刺激后,MMP-20的基因表达上调(P<0.05),MMP-20启动子在(-157～+23)之前的区域转录活性增强(P<0.05);EGFR、ErbB2、ERK阻断剂均能抑制EGF对MMP-20基因表达的调控(P<0.05);转录因子C-Jun介导EGFR/ErbB2上调MMP-20基因的表达(P<0.05);突变MMP-20启动子的AP1结合位点后,MMP-20基因表达的水平下降(P<0.05),同时EGF/ErbB2也失去上调MMP-20基因表达水平的作用。结论:在成釉细胞中EGF通过EGFR/ErbB2～ERK～C-Jun信号通路对MMP-20基因的表达起到调控作用。
AIM： To investigate the roles of EGF/EGFR/ErbB2 signaling pathway in the regulation of ma- trix metalloproteinase-20 （MMP-20） expression in mouse ameloblasts. METHODS ： Mouse ameloblasts were stimula- ted with 20 ng/mL EGF. MMP-20 expression in ameloblasts was detected by RT-PCR. Changes of MMP-20 promoter was analyzed by dual luciferase reporter analysis. The effects of the inhibitors of EGFR, ErbB2, MAPK signaling path- way and c - Jun pathway on MMP-20 promoter transcription activity were investigated by dual luceferase report gene a- nalysis. RESULTS： EGF enhanced MMP-20 expression and MMP-20 promoter activity before （ - 157 - ＋ 23 ）region in ameloblasts （P 〈 0.05）. EGFR, ErbB2 and ERK inhibitors attenuated the effects of EGF on MMP-20 expression. C-Jun increased MMP-20 expression through EGFR/ErbB2. Mutation of MMP-20 promoter at AP1 binding site re- duced MMP- 20 expression. CONCLUSION： EGF can induce MMP-20 expression via EGFR/ErbB2-ERK-C-Junsignaling pathway in mouse ameloblasts.
出处
《牙体牙髓牙周病学杂志》
CAS
北大核心
2013年第7期417-423,共7页
Chinese Journal of Conservative Dentistry
基金
国家自然科学基金(30973327)
山东省自然科学基金(ZR2010HM076)
