The effect of TRPM7 suppression on the proliferation, migration and osteogenic differentiation of human dental pulp stem cells
The effect of TRPM7 suppression on the proliferation, migration and osteogenic differentiation of human dental pulp stem cells
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DOI：
10.1111/iej.12193
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发表时间：
2014-06-01
影响因子：
5
通讯作者：
Wu, B. L.
中科院分区：
文献类型：
--
作者：
Cui, L.;Xu, S. M.;Wu, B. L.
目的探讨钙镁离子通道TRPM 7在人牙髓干细胞（human dental pulp stem cells，hDPSCs）增殖、迁移和成骨分化中的作用。用表达TRPM 7特异性短发夹RNA（shRNA）或对照shRNA的重组慢病毒感染分离的hDPSC，以抑制TRPM 7 mRNA表达并研究其功能作用。使用MTT测定法测量活细胞数和细胞周期分析来评估shRNA感染的hDPSC的增殖。使用transwell测定评估细胞迁移。结果TRPM 7在人牙髓干细胞中广泛表达，主要表达于细胞膜和细胞质中，并与对照组相比差异有统计学意义（P < 0. 05）。TRPM 7的抑制抑制了hDPSC的增殖和迁移能力。TRPM 7 mRNA表达在hDPSC成骨分化过程中升高。TRPM 7特异性shRNA抑制hDPSC的成骨分化，并下调成骨标志物碱性磷酸酶（ALP）、牙本质涎磷蛋白（DSPP）、骨涎蛋白（BSP）、Runt相关转录因子（RUNX 2）和osterix（OSX）的mRNA表达。
AimTo investigate the role of the Ca2+-Mg2+ ion channel TRPM7 in the proliferation, migration and osteogenic differentiation of human dental pulp stem cells (hDPSCs).MethodologyImmunohistochemistry was used to localize expression of TRPM7 in human dental pulp tissues and in cultured hDPSCs. Isolated hDPSCs were infected with recombinant lentiviruses expressing short hairpin RNA (shRNA) specific for TRPM7, or control shRNA, in order to suppress TRPM7 mRNA expression and investigate its functional role. The proliferation of the shRNA-infected hDPSCs was evaluated using both an MTT assay to measure viable cell numbers and cell cycle analysis. Cell migration was assessed using a transwell assay. The dynamic mRNA expression of TRPM7 during osteogenic differentiation of hDPSCs and the effect of shRNA specific for TRPM7 on hDPSC osteogenic differentiation were evaluated by real-time PCR.ResultsTRPM7 expression was widespread in human dental pulp tissue and was detected mainly in the cytomembrane and cytoplasm of hDPSCs. Suppression of TRPM7 inhibited both the proliferation and the migratory capacity of hDPSCs. TRPM7 mRNA expression was elevated during osteogenic differentiation of hDPSCs. TRPM7-specific shRNA inhibited osteogenic differentiation of hDPSCs, with downregulated mRNA expression of the osteogenic markers alkaline phosphatase (ALP), dentine sialophosphoprotein (DSPP), bone sialoprotein (BSP), runt-related transcription factor (RUNX2) and osterix (OSX).ConclusionsTRPM7 was involved in the regulation of hDPSC proliferation, migration and osteogenic differentiation and may play a role in the dental pulp repair process.