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The effect of TRPM 7 suppression on the proliferation, migration and osteogenic differentiation of human dental pulp stem cells
Author(s) -
Cui L.,
Xu S. M.,
Ma D. D.,
Wu B. L.
Publication year - 2014
Publication title -
international endodontic journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.988
H-Index - 119
eISSN - 1365-2591
pISSN - 0143-2885
DOI - 10.1111/iej.12193
Subject(s) - small hairpin rna , dental pulp stem cells , trpm7 , microbiology and biotechnology , bone sialoprotein , runx2 , chemistry , transcription factor , biology , mesenchymal stem cell , alkaline phosphatase , osteocalcin , rna , biochemistry , receptor , transient receptor potential channel , gene , enzyme
Aim To investigate the role of the Ca 2+ –Mg 2+ ion channel TRPM7 in the proliferation, migration and osteogenic differentiation of human dental pulp stem cells ( hDPSC s). Methodology Immunohistochemistry was used to localize expression of TRPM7 in human dental pulp tissues and in cultured hDPSC s. Isolated hDPSC s 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 hDPSC s 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 hDPSC s and the effect of shRNA specific for TRPM7 on hDPSC osteogenic differentiation were evaluated by real‐time PCR. Results TRPM7 expression was widespread in human dental pulp tissue and was detected mainly in the cytomembrane and cytoplasm of hDPSC s. Suppression of TRPM7 inhibited both the proliferation and the migratory capacity of hDPSC s. TRPM7 mRNA expression was elevated during osteogenic differentiation of hDPSC s. TRPM7‐specific shRNA inhibited osteogenic differentiation of hDPSC s, 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). Conclusions TRPM7 was involved in the regulation of hDPSC proliferation, migration and osteogenic differentiation and may play a role in the dental pulp repair process.