MAPK-ERK在牙本质基质蛋白1信号通路中的作用  

DMP1's signal pathway via MAPK-ERK

作　　者：伍虹[1] 庄沛林[1] 余艳崧[1] 陈绛媛[1] Sfeir Charles 黄洪章[3] 

机构地区：[1]中山大学孙逸仙纪念医院口腔科,广州510120 [2]美国匹兹堡大学牙学院·颅颌面重建中心 [3]中山大学光华口腔医学院·附属口腔医院,广东省口腔医学重点实验室

出　　处：《中华口腔医学研究杂志（电子版）》2013年第6期1-5,共5页Chinese Journal of Stomatological Research(Electronic Edition)

基　　金：国家自然科学基金(81200825);广东省医学科研基金(A2011176)

摘　　要：目的探讨在人骨髓间充质干细胞(hMSC)、小鼠前成骨细胞(MC3T3)和成牙本质细胞(MDPC-23)中,牙本质基质蛋白1(DMP1)是否通过激活丝裂原活化蛋白激酶(MAPK)-细胞外调节蛋白激酶(ERK)信号通路发挥作用。方法 Western blot检测不同时间点rDMP1C和rDMP1F蛋白处理hMSC、MC3T3-E1和MDPC-23后,对MAPK-ERK的激活情况;并检测使用MAPK抑制剂后,ERK的激活是否被抑制;细胞免疫荧光技术观察MAPK抑制剂抑制激活的ERK从胞浆向胞核的转位情况。Western blot检测siRNA沉默Ras基因后,对rDMP1引起MAPK-ERK信号通路激活的影响。结果三种细胞中,rDMP1F和rDMP1C在5 min^3 h均可以激活MAPK-ERK通路,而总ERK在各时间点均无显著变化;rDMP1F激活信号通路的持续时间均明显长于rDMP1C;MAPK抑制剂处理组的p-ERK条带与rDMP1F/C处理组的p-ERK条带差别明显。hMSC中,rDMP1F激活MAPK信号通路持续时间要长于其在MC3T3和MDPC-23中的作用。细胞免疫荧光检测发现,MAPK抑制剂组可以阻断ERK向细胞核内的转位。MC3T3和MDPC-23在siRNA沉默Ras基因后,ERK的磷酸化水平与rDMP1F单独处理组相比显著降低。结论 rDMP1C和rDMP1F都通过Ras激活MAPK-ERK信号通路,而rDMP1F比rDMP1C具有更强的信号功能。Objective Dentin matrix phosphoprotein 1 （DMP1） is a non-collagenous, acidic extracellular matrix protein expressed chiefly in bone and dentin. The present study was aimed to investigate whether DMP1 can trigger MAPK signalling pathway in hMSC, MC3T3-E1 and MDPC-23 cells. Methods The activation of MAPK-ERK by rDMP1F/C and inhibition of ERK by MAPK inhibitor at different timepoints in hMSC, MC3T3-E1 and MDPC-23 cells was detected using Western blot. The inhibition of ERK cytoplasm/nucleus translocation by the MAPK inhibitor was detected using immunofluorescence microscopy. The activation of MAPK-ERK signalling pathway by DMP1 was further validated through knowdown of Ras gene expression by siRNA. Results The MAPK-ERK could be activated by both recombinant DMP1 C-terminal （rDMP1C） and full lenth （rDMP1F） from 5 min to 3 h in three cell lines. The phosphorylated-ERK increased, but not total ERK, in these cell lines treated by rDMP1. The phosphorylation of MAPK-ERK by rDMP1F in hMSC cells last longer than that in MC3T3 or MDPC-23 cells. The activation of MAPK lasted longer by rDMP1F than by rDMP1C. MAPK inhibitor could decrease the phosphorylation of MAPK-ERK by rDMP1C and rDMP1F and inhibit the ERKtranslocation from the cytoplasm into the nucleus. Knowdown of Ras gene expression by siRNA could significantly decrease the phosphoralated of ERK. Conclusion Both rDMP1C and rDMP1F can active the MEK-ERK, however, rDMP1F has a stronger effect in trigger MAPK signalling pathway than rDMP1C.

关 键 词：牙本质基质蛋白1 丝裂原活化蛋白激酶 细胞外调节蛋白激酶 信号通路 

分 类 号：R780.2[医药卫生—口腔医学]