title: SRC kinase family inhibitor PP2 promotes DMSO-induced cardiac differentiation of P19 cells and inhibits proliferation.
abstract: BACKGROUND
  It has been reported recently that PP2, a Src family kinase inhibitor, promotes selective cardiogenesis in embryonic stem cells. However, there is no other research proved pro-cardiogenic characteristic of PP2 so far. In this study, we explored the potential cardiogenic effect of PP2 on P19 cells differentiation.
  
  
  METHODS
  P19-αMHC-EGFP cell line was established by transfecting P19 cells with αMHC-EGFP vector in order to evaluate cardiogenesis with EGFP. P19-αMHC-EGFP cells and P19 cells were induced to differentiate into cardiomyocytes with 1%DMSO, 5 μmol/L PP2, or both 1%DMSO and 5 μmol/L PP2. Differentiated cells from P19-αMHC-EGFP cells were then assessed under confocal microscope. Western-blot and RT-PCR were also performed to detect expression of cardiac troponin I and cardiac transcription factors respectively. In addition, the effects of PP2 on proliferation of P19 cells were further examined using Cell Counting Kit-8.
  
  
  RESULTS
  EGFP positive cells were firstly detected on day 7 and PP2 alone cannot induce efficient cardiac differentiation of P19-αMHC-EGFP cells. However PP2 supplementation dramatically increases DMSO induced cardiac differentiation than DMSO alone. It was also found that PP2 inhibit proliferation of P19 cells in both a dose-dependent manner and a time-dependent manner.
  
  
  CONCLUSIONS
  PP2 alone cannot substitute DMSO to induce cardiac differentiation, however, PP2 supplementation drastically promotes DMSO-induced cardiac differentiation of P19 cells. The increased percentages of differentiated cardiac myocytes is partly resulting from cell proliferative inhibit effect of PP2 in undifferentiated P19 cells. P19-αMHC-EGFP cell line has the potential to be used for regenerative therapies in experimental models of heart repair.
authors.affiliation: Division of Cardiology, Department of Medicine, The Affiliated Hospital of Jiangsu University, Zhenjiang, PR China 212001. gongjnjmu@hotmail.com
authors.forename: Jie
authors.initials: J
authors.lastname: Gong
authors.forename: Hai-yong
authors.initials: HY
authors.lastname: Gu
authors.forename: Xiao
authors.initials: X
authors.lastname: Wang
authors.forename: Yi
authors.initials: Y
authors.lastname: Liang
authors.forename: Tao
authors.initials: T
authors.lastname: Sun
authors.forename: Pei-jing
authors.initials: PJ
authors.lastname: Liu
authors.forename: Yi
authors.initials: Y
authors.lastname: Wang
authors.forename: Jin-chuan
authors.initials: JC
authors.lastname: Yan
authors.forename: Zhi-jun
authors.initials: ZJ
authors.lastname: Jiao
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D004121:Dimethyl Sulfoxide / Q000008:administration & dosage*; D004357:Drug Synergism; D051379:Mice; D008809:Mice, Inbred C3H; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000201:enzymology; D011743:Pyrimidines / Q000008:administration & dosage*; D019061:src-Family Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: C412373:AG 1879; D011743:Pyrimidines; D019061:src-Family Kinases; D004121:Dimethyl Sulfoxide