@Article{biocell.2023.029080,
AUTHOR = {YANG CAO, YINING LIU, YANG YU, XIAOFEI GUO, XIUXIU WANG, WENYA MA, HANJING LI, ZHONGYU REN, XINLU GAO, SIJIA LI, HAOYU JI, HONGYANG CHEN, HONG YAN, YANAN TIAN, XIN WANG, BENZHI CAI},
TITLE = {LncRNA <i>ZFAS1</i> regulates cardiomyocyte differentiation of human embryonic stem cells},
JOURNAL = {BIOCELL},
VOLUME = {47},
YEAR = {2023},
NUMBER = {6},
PAGES = {1407--1416},
URL = {http://www.techscience.com/biocell/v47n6/52790},
ISSN = {1667-5746},
ABSTRACT = {<b>Background:</b> Cardiomyocytes derived from human embryonic stem cells (hESCs) are regulated by complex and stringent gene networks during differentiation. Long non-coding RNAs (lncRNAs) exert critical epigenetic regulatory functions in multiple differentiation processes. However, the involvement of lncRNAs in the differentiation of hESCs into cardiomyocytes has not yet been fully elucidated. Here, we identified the key roles of <i>ZFAS1</i> (lncRNA zinc finger antisense 1) in the differentiation of cardiomyocytes from hESCs. <b>Methods:</b> A model of cardiomyocyte differentiation from stem cells was established using the monolayer differentiation method, and the number of beating hESCs-derived cardiomyocytes was calculated. Gene expression was analyzed by quantitative real-time PCR (qRT-PCR). Immunofluorescence assays were performed to assess the expression of cardiac troponin T (cTnT) and α-actinin protein in cardiomyocytes. <b>Results:</b> qRT-PCR showed that <i>ZFAS1</i> expression in the mesoderm was significantly higher than that in embryonic stem cells, cardiac progenitor cells, and cardiomyocytes. Knockdown of <i>ZFAS1</i> inhibited cardiomyocyte differentiation from hESCs, which was characterized by reduced expression of the cardiac-specific markers cTnT, α-actinin, myosin heavy chain 6 (MYH6), and myosin heavy chain 7 (MYH7). In contrast, <i>ZFAS1</i> overexpression remarkably increased the percentage of spontaneously beating cardiomyocytes. In terms of the mechanism, we found that <i>ZFAS1</i> is an antisense lncRNA at the 5′ end of the protein-coding gene <i>ZNFX1</i>. Knockdown of <i>ZFAS1</i> could increase the mRNA expression level of <i>ZNFX1</i>. Furthermore, qRT-PCR demonstrated that the silencing of <i>ZNFX1</i> led to an increase in cardiac-specific markers that predicted the promotion of cardiomyocyte differentiation. <b>Conclusion:</b> Altogether, these data suggest that lncRNA-<i>ZFAS1</i> is required for cardiac differentiation by functionally inhibiting the expression of <i>ZNFX1</i>, which may provide a reference for the treatment of heart disease to a certain extent.},
DOI = {10.32604/biocell.2023.029080}
}