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Interaction between mD ia1 and ROCK in Rho‐induced migration and adhesion of human dental pulp cells
Author(s) -
Cheng L.,
Xu J.,
Qian Y. Y.,
Pan H. Y.,
Yang H.,
Shao M. Y.,
Cheng R.,
Hu T.
Publication year - 2017
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.12587
Subject(s) - focal adhesion , paxillin , cell migration , microbiology and biotechnology , cell adhesion , rhoa , rho associated protein kinase , chemistry , adhesion , phosphorylation , biology , biochemistry , cell , signal transduction , organic chemistry
Aim To investigate the effects of mammalian homologue of Drosophila diaphanous‐1( mD ia1) and Rho‐associated coiled‐coil‐containing protein kinase ( ROCK ) on the migration and adhesion of dental pulp cells ( DPC s). Methodology Lysophosphatidic acid ( LPA ) was used to activate Rho signalling. mD ia1 and ROCK were inhibited by short interfering RNA and the specific inhibitor, Y‐27632, respectively. The migration of DPC s was assessed using the transwell migration assay and scratch test. Formation of cytoskeleton and focal adhesions(FAs) was observed by confocal laser scanning microscopy. Cell adhesion and spreading assays were performed. Phosphorylation of focal adhesion kinase ( FAK ) and paxillin was detected by Western blotting, and the bands were analysed using Adobe Photoshop CS 5 software. All experiments were performed at least three times, and data were analysed with one‐way anova and a post hoc test. Results LPA ‐triggered activation of Rho and inhibition of ROCK significantly increased the cell migration rate. Cell migration was inhibited by silencing mD ia1. mD ia1 silencing and ROCK inhibition suppressed the LPA ‐induced formation of the cytoskeleton, FA and phosphorylation of FAK and paxillin. Inhibition of ROCK or mD ia1 facilitated early cell adhesion and spreading; by contrast, the combined inhibition of ROCK and mD ia1 neutralized these effects. Conclusions mD ia1 promoted RhoA‐induced migration of DPC s, but ROCK had an opposite effect. Both mD ia1 and ROCK participated in cytoskeleton formation and adhesion of DPC s. The interactions between mD ia1 and ROCK might influence dental pulp repair by determining the migration and adhesion of DPCs.