title: Regulation of c-kit+ progenitor cells by stromal cell derived factor-1α in adult murine heart.
abstract: BACKGROUND
  c-kit-positive cardiac progenitor cells (CPCs) have been proven suitable for stem cell therapy. CPCs marker c-kit and its ligand, the stem cell factor (SCF), are 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.
  
  
  RESULTS
  CPCs were isolated from adult mouse hearts, and c-kit-positive CPCs were purified by magnetic-activated c-kit cell sorting magnetic beads. The cells were cultured with SDF-1α, c-kit expression was measured by western blotting and qPCR, the proliferation and migration of cells were measured by CCK-8 and transwell assay, DNA methyltransferase (DNMT) mRNA were measured by qPCR, global DNMT activity was measured by DNMT activity assay kit, and DNA methylation was analysed using Sequenom's MassARRAY platform. Results showed that SDF-1α could enhance the expression of c-kit, which results in the promoting of c-kit-positive CPCs proliferation and migration. SDF-1α stimulation inhibited the expression of DNMT1, DNMT3β, and global DNMT activity, which led to significant demethylation in c-kit-positive CPCs.
  
  
  CONCLUSIONS
  SDF-1α signalling, via CXCR4 activation, up-regulated c-kit expression by inhibiting DNMT1 and DNMT3β expression and global DNMT activity, and by subsequent demethylation of the c-kit gene.
authors.affiliation: Department of Cardiology, Zhongda Hospital, Medical School of Southeast University, Nanjing, Jiangsu 210009, China.
authors.forename: Zhongpu
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Cardiology, Zhongda Hospital, Medical School of Southeast University, Nanjing, Jiangsu 210009, China.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Pan
authors.affiliation: Department of Cardiology, Zhongda Hospital, Medical School of Southeast University, Nanjing, Jiangsu 210009, China.
authors.forename: Yuyu
authors.initials: Y
authors.lastname: Yao
authors.affiliation: Department of Cardiology, Zhongda Hospital, Medical School of Southeast University, Nanjing, Jiangsu 210009, China.
authors.forename: Fengdi
authors.initials: F
authors.lastname: Yan
authors.affiliation: Department of Cardiology, Zhongda Hospital, Medical School of Southeast University, Nanjing, Jiangsu 210009, China.
authors.forename: Long
authors.initials: L
authors.lastname: Chen
authors.affiliation: Department of Cardiology, Zhongda Hospital, Medical School of Southeast University, Nanjing, Jiangsu 210009, China.
authors.forename: Rong
authors.initials: R
authors.lastname: Huang
authors.affiliation: Department of Cardiology, Zhongda Hospital, Medical School of Southeast University, Nanjing, Jiangsu 210009, China. Electronic address: magenshan@hotmail.com.
authors.forename: Genshan
authors.initials: G
authors.lastname: Ma
mesh_terms: D000818:Animals; D002465:Cell Movement / Q000502:physiology; D049109:Cell Proliferation; D054377:Chemokine CXCL12 / Q000378:metabolism*; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000378:metabolism; D019175:DNA Methylation / Q000502:physiology; D005786:Gene Expression Regulation / Q000502:physiology*; D051379:Mice; D009206:Myocardium / Q000166:cytology / Q000378:metabolism*; D019009:Proto-Oncogene Proteins c-kit / Q000096:biosynthesis*; D015398:Signal Transduction / Q000502:physiology*; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism*; D000097584:DNA Methyltransferase 3B
chemical_list: D054377:Chemokine CXCL12; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; D019009:Proto-Oncogene Proteins c-kit; D000097584:DNA Methyltransferase 3B; C515256:Cxcl12 protein, mouse; C000619381:Dnmt1 protein, mouse
keywords: CXCR4; Cardiac progenitor cells; DNA methylation; DNA methyltransferase; Stromal cell derived factor-1α; c-kit