title: Downregulation of long non-coding RNA H19 promotes P19CL6 cells proliferation and inhibits apoptosis during late-stage cardiac differentiation via miR-19b-modulated Sox6.
abstract: BACKGROUND
  Regulating cardiac differentiation to maintain normal heart development and function is very important. At present, biological functions of H19 in cardiac differentiation is not completely clear.
  
  
  METHODS
  To explore the functional effect of H19 during cardiac differentiation. Expression levels of early cardiac-specific markers Nkx-2.5 and GATA4, cardiac contractile protein genes α-MHC and MLC-2v were determined by qRT-PCR and western lot. The levels of lncRNA H19 and miR-19b were detected by qRT-PCR. We further predicted the binding sequence of H19 and miR-19b by online softwares starBase v2.0 and TargetScan. The biological functions of H19 and Sox6 were evaluated by CCK-8 kit, cell cycle and apoptosis assay and caspase-3 activity.
  
  
  RESULTS
  The expression levels of α-MHC, MLC-2v and H19 were upregulated, and miR-19b was downregulated significantly in mouse P19CL6 cells at the late stage of cardiac differentiation. Biological function analysis showed that knockdown of H19 promoted cell proliferation and inhibits cell apoptosis. H19 suppressed miR-19b expression and miR-19b targeted Sox6, which inhibited cell proliferation and promoted apoptosis in P19CL6 cells during late-stage cardiac differentiation. Importantly, Sox6 overexpression could reverse the positive effects of H19 knockdown on P19CL6 cells.
  
  
  CONCLUSIONS
  Downregulation of H19 promoted cell proliferation and inhibited cell apoptosis during late-stage cardiac differentiation by regulating the negative role of miR-19b in Sox6 expression, which suggested that the manipulation of H19 expression could serve as a potential strategy for heart disease.
authors.affiliation: Children's Heart Center, Henan Province People's Hospital, Zhengzhou, 450003 China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Han
authors.affiliation: Department of Cardiology, Henan Province People's Hospital, No. 7 Weiwu Road, Zhengzhou, 450003 China.
authors.forename: Hongdang
authors.initials: H
authors.lastname: Xu
authors.affiliation: Department of Cardiology, Henan Province People's Hospital, No. 7 Weiwu Road, Zhengzhou, 450003 China.
authors.forename: Jiangtao
authors.initials: J
authors.lastname: Cheng
authors.affiliation: Children's Heart Center, Henan Province People's Hospital, Zhengzhou, 450003 China.
authors.forename: Yanwei
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Cardiology, Henan Province People's Hospital, No. 7 Weiwu Road, Zhengzhou, 450003 China.
authors.forename: Chuanyu
authors.initials: C
authors.lastname: Gao
authors.affiliation: Children's Heart Center, Henan Province People's Hospital, Zhengzhou, 450003 China.
authors.forename: Taibing
authors.initials: T
authors.lastname: Fan
authors.affiliation: Children's Heart Center, Henan Province People's Hospital, Zhengzhou, 450003 China.
authors.forename: Bangtian
authors.initials: B
authors.lastname: Peng
authors.affiliation: Children's Heart Center, Henan Province People's Hospital, Zhengzhou, 450003 China.
authors.forename: Bin
authors.initials: B
authors.lastname: Li
authors.affiliation: Department of Ultrasonography, Henan Province People's Hospital, Zhengzhou, 450003 China.
authors.forename: Lin
authors.initials: L
authors.lastname: Liu
authors.affiliation: Department of Cardiovascular Surgery, Henan Province People's Hospital, Zhengzhou, 450003 China.
authors.forename: Zhaoyun
authors.initials: Z
authors.lastname: Cheng
keywords: Cardiac differentiation; H19; IncRNA; Sox6; miR-19b