Knockdown of nucleosome assembly protein 1-like 1 promotes dimethyl sulfoxide-induced differentiation of P19CL6 cells into cardiomyocytes.
Transplantation of cardiomyocytes derived from stem cells is a promising option for cardiac repair. However, how to obtain efficient cardiomyocytes from stem cells is still a great challenge. Understanding of the mechanism that regulates the cardiac differentiation of stem cells is necessary for the effective induction of cardiomyocytes. A clonal derivative named P19CL6 cells can easily differentiate into cardiomyocytes with 1% dimethyl sulfoxide (DMSO) treatment, which offers a valuable model to study cardiomyocytes differentiation in vitro. In this study, the isobaric tags for relative and absolute quantitation (iTRAQ) proteomics were performed to identify proteins associated with cardiomyocytes differentiation of P19CL6 cells induced by DMSO. Out of 543 non-redundant proteins identified, 207 proteins showed significant changes during differentiation with ≥1.2-fold or ≤0.83-fold changes cut-offs. Nine proteins were confirmed by the quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot analysis respectively. Notably, broad consistency was well showed between mRNA and protein expression for down-regulation of nucleosome assembly protein 1-like 1 (Nap1l1). Further study revealed that knockdown of Nap1l1 by stable transfection of shRNA vector significantly accelerated DMSO-induced cardiomyocytes differentiation of P19CL6 cells characterized by increases in expression of cardiac specific transcription factors, genes, and proteins (GATA4, MEF-2C, ANP, BNP, cTNT, and β-MHC). Therefore, Nap1l1 is a novel protein that regulates cardiomyocytes differentiation of P19CL6 cells induced by DMSO.
Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China.
Lu
L
Li
Hui
H
Gong
Hongxiu
H
Yu
Xiaohui
X
Liu
Qingping
Q
Liu
Guoquan
G
Yan
Yang
Y
Zhang
Haojie
H
Lu
Yunzeng
Y
Zou
Pengyuan
P
Yang
D000818:Animals; D014408:Biomarkers, Tumor / Q000032:analysis / Q000378:metabolism; D015153:Blotting, Western; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D054278:Embryonal Carcinoma Stem Cells / Q000187:drug effects* / Q000378:metabolism; D050991:GATA4 Transcription Factor / Q000235:genetics / Q000378:metabolism; D055785:Gene Knockdown Techniques / Q000379:methods*; D051379:Mice; D032383:Myocytes, Cardiac; D056490:Nucleosome Assembly Protein 1 / Q000235:genetics / Q000378:metabolism*; D020543:Proteome / Q000032:analysis; D040901:Proteomics; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D013194:Staining and Labeling; D014162:Transfection
D014408:Biomarkers, Tumor; D050991:GATA4 Transcription Factor; C484171:Gata4 protein, mouse; C533558:Nap1l1 protein, mouse; D056490:Nucleosome Assembly Protein 1; D020543:Proteome; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D004121:Dimethyl Sulfoxide