Effects of verapamil on the immediate-early gene expression of bone marrow mesenchymal stem cells stimulated by mechanical strain in vitro
Runguang Li, Mingfa Wei, Jingfan Shao
Med Sci Monit Basic Res 2013; 19:68-75
To study the effects of verapamil on the immediate-early genes (IEGs) expression of bone marrow mesenchymal stem cells (MSCs) stimulated by cyclic mechanical strain, in order to deduce the role of calcium ion channel in the cell signaling responses of MSCs to mechanical strain.
Material and Methods: MSCs were isolated and cultured, and the passage of 3–6 MSCs were stimulated by mechanical strain and pretreated with or without verapamil. After that, flow cytometry was used to measure the fluorescence intensity of intracellular Ca2+ immediately. The expression of early-response genes/proteins (c-fos, c-jun and c-myc) were examined by RT-PCR, immunohistochemistry and Western blot.
Results: Intracellular Ca2+ concentration of MSCs significantly changed when stimulated by cyclic strain, and the expression of c-fos, c-jun and c-myc remarkably increased in both mRNA and protein levels, while verapamil pre-treatment partially inhibited these effects (P<0.01).
Conclusions: The changes of the intracellular calcium concentration of MSCs induced by mechanical strain, dependent on the regulation of calcium channel activation, might play a role in the early response of MSCs to cyclic strain.
Keywords: Proto-Oncogene Proteins c-jun - metabolism, Proto-Oncogene Proteins c-fos - metabolism, Mesenchymal Stromal Cells - drug effects, Ions, Ion Channels - metabolism, Genes, Immediate-Early, Gene Expression Regulation - drug effects, Child, Preschool, Child, Cells, Cultured, Calcium Channel Blockers - pharmacology, Calcium - metabolism, Proto-Oncogene Proteins c-myc - metabolism, Signal Transduction, Stress, Mechanical, Verapamil - pharmacology