Effects of human full-length amelogenin on the proliferation of human mesenchymal stem cells derived from bone marrow
Effects of human full-length amelogenin on the proliferation of human mesenchymal stem cells derived from bone marrow
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DOI：
10.1007/s00441-010-1064-7
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发表时间：
2010-11-01
影响因子：
3.6
通讯作者：
Tanne, Kazuo
中科院分区：
文献类型：
--
作者：
Huang, Yu-Ching;Tanimoto, Kotaro;Tanne, Kazuo
釉原蛋白是在釉质形成中起关键作用的釉质基质蛋白。最近的研究表明，釉原蛋白还具有细胞信号传导特性。虽然釉原蛋白被认为是成釉细胞的特异性产物，但最近的研究表明其在骨髓基质细胞中表达。在这项研究中，我们研究了重组人全长釉原蛋白（rh 174）对人骨髓间充质干细胞（MSCs）增殖的影响，并表征了细胞内信号通路的相关变化。用剂量范围为0至1，000 ng/ml的rh 174处理MSC。用溴脱氧尿苷（BrdU）免疫分析法检测细胞增殖活性。分析了MSC中溶酶体相关膜蛋白1（LAMP 1）的表达，LAMP 1可能是一种釉原蛋白受体。使用抗LAMP 1抗体阻断rh 174与LAMP 1的结合。采用细胞信号激活ELISA（CASE）试剂盒和western blot分析检测MAPK-ERK通路。使用特异性MAPK抑制剂U 0126来阻断ERK活性。结果表明rh 174可促进MSCs的增殖和MAPK-ERK活性。加入抗LAMP 1抗体可抑制rh 174促进的MSC增殖和MAPK-ERK活性。U 0126可抑制rh 174诱导的MSCs增殖。总之，它表明，rh 174通过MAPK-ERK信号通路与LAMP I相互作用，增加MSC的增殖，表明MSC应用于组织再生的可能性在口面部区域。
Amelogenins are enamel matrix proteins that play a crucial role in enamel formation. Recent studies have revealed that amelogenins also have cell signaling properties. Although amelogenins had been described as specific products of ameloblasts, recent research has demonstrated their expression in bone marrow stromal cells. In this study, we examined the effect of recombinant human full-length amelogenin (rh174) on the proliferation of human mesenchymal stem cells (MSCs) derived from bone marrow and characterized the associated changes in intracellular signaling pathways. MSCs were treated with rh174 ranging in dose from 0 to 1,000 ng/ml. Cell proliferative activity was analyzed by bromodeoxyuridine (BrdU) immunoassay. The expression of lysosomal-associated membrane protein 1 (LAMP 1), a possible amelogenin receptor, in MSCs was analyzed. Anti-LAMP 1 antibody was used to block the binding of rh174 to LAMP1. The MAPK-ERK pathway was examined by Cellular Activation of Signaling ELISA (CASE) kit and western blot analysis. A specific MAPK inhibitor, U0126, was used to block ERK activity. It was shown that rh174 increased the proliferation of MSCs and MAPK-ERK activity. The MSC proliferation and MAPK-ERK activity enhanced by rh174 were reduced by the addition of anti-LAMP1 antibody. Additionally, the increased proliferation of MSCs induced by rh174 was inhibited in the presence of U0126. In conclusion, it is demonstrated that rh174 increases the proliferation of MSCs by interaction with LAMP I through the MAPK-ERK signaling pathway, indicating the possibility of MSC application to tissue regeneration in the orofacial region.