Abstract
Cardiac progenitor cells (CPCs) have been proven suitable for stem cell therapy after myocardial infarction, especially c-kit(+)CPCs. CPCs marker c-kit and its ligand, the stem cell factor (SCF), are linked as c-kit/SCF axis, which is associated with the functions of proliferation and differentiation. In our previous study, we found that stromal cell-derived factor-1α (SDF-1α) could enhance the expression of c-kit. However, the mechanism is unknown. CPCs were isolated from adult mouse hearts, c-kit(+) and c-kit(-) CPCs were separated by magnetic beads. The cells were cultured with SDF-1α and CXCR4-selective antagonist AMD3100, and c-kit expression was measured by qPCR and Western blotting. Results showed that SDF-1α could enhance c-kit expression of c-kit(+)CPCs, made c-kit(-)CPCs expressing c-kit, and AMD3100 could inhibit the function of SDF-1α. After the intervention of SDF-1α and AMD3100, proliferation and migration of CPCs were measured by CCK-8 and transwell assay. Results showed that SDF-1α could enhance the proliferation and migration of both c-kit(+) and c-kit(-) CPCs, and AMD3100 could inhibit these functions. DNA methyltransferase (DNMT) mRNA were measured by qPCR, DNMT activity was measured using the DNMT activity assay kit, and DNA methylation was analyzed using Sequenom's MassARRAY platform, after the CPCs were cultured with SDF-1α. The results showed that SDF-1α stimulation inhibited the expression of DNMT1 and DNMT3β, which are critical for the maintenance of regional DNA methylation. Global DNMT activity was also inhibited by SDF-1α. Lastly, SDF-1α treatment led to significant demethylation in both c-kit(+) and c-kit(-) CPCs. SDF-1α combined with CXCR4 could up-regulate c-kit expression of c-kit(+)CPCs and make c-kit(-)CPCs expressing c-kit, which result in the CPCs proliferation and migration ability improvement, through the inhibition of DNMT1 and DNMT3β expression and global DNMT activity, as well as the subsequent demethylation of the c-kit gene.
Highlights
Ischemic heart disease remain the leading causes of mortality and morbidity worldwide, and stem cell therapy may regenerate cardiac tissue directly by inducing neovasculogenesis and cardiogenesis
stromal cell-derived factor-1a (SDF-1a) combined with CXCR4 could up-regulate c-kit expression of c-kit(+)Cardiac progenitor cells (CPCs) and make c-kit(2)CPCs expressing c-kit, which result in the CPCs proliferation and migration ability improvement, through the inhibition of DNMT1 and DNMT3b expression and global DNA methyltransferase (DNMT) activity, as well as the subsequent demethylation of the c-kit gene
The present study demonstrates that SDF-1a combined with CXCR4 could up-regulate c-kit expression of c-kit(+)CPCs and make c-kit(2)CPCs expressing c-kit, which result in the CPCs proliferation and migration ability improvement, through the inhibition of DNMT1 and DNMT3b expression and global DNMT activity, as well as the subsequent demethylation of the ckit gene
Summary
Ischemic heart disease remain the leading causes of mortality and morbidity worldwide, and stem cell therapy may regenerate cardiac tissue directly by inducing neovasculogenesis and cardiogenesis. The heart have several populations of CPCs, which are self-renewing, clonogenic, multipotent and have the ability to proliferate and differentiate into functional cardiomyocytes, smooth muscle cells, and other kinds of cells [2,4,5,6]. Among these CPCs, c-kit(+)CPCs are especially suitable in cell therapy for the recovery of injured cardiomyocytes [2,4]. Cardiac progenitor cells (CPCs) have been proven suitable for stem cell therapy after myocardial infarction, especially c-kit(+)CPCs. CPCs marker c-kit and its ligand, the stem cell factor (SCF), are linked as c-kit/SCF axis, which is associated with the functions of proliferation and differentiation.
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