LncRNA-uc.40 silence promotes P19 embryonic cells differentiation to cardiomyocyte via the PBX1 gene

Abstract

Uc.40 is a long noncoding RNA that is highly conserved among different species, although its function is unknown. It is highly expressed in abnormal human embryonic heart. We previously reported that overexpression of uc.40 promoted apoptosis and inhibited proliferation of P19 cells, and downregulated PBX1, which was identified as a potential target gene of uc.40. The current study evaluated the effects of uc40-siRNA-44 (siRNA against uc.40) on the differentiation, proliferation, apoptosis, and mitochondrial function in P19 cells, and investigated the relationship between uc.40 and PBX1 in cardiomyocytes. The uc.40 silencing expression was confirmed by quantitative real-time polymerase chain reaction (RT-PCR). Observation of morphological changes in transfected P19 cells during different stages of differentiation revealed that uc40-siRNA-44 increased the number of cardiomyocyes. There was no significant difference in the morphology or time of differentiation between the uc40-siRNA-44 group and the control group. uc40-siRNA-44 significantly promoted proliferation of P19 cells and inhibited serum starvation-induced apoptosis. There was no significant difference in mitochondrial DNA copy number or cellular ATP level between the two groups, and ROS levels were significantly decreased in uc40-siRNA-44-transfected cells. The levels of PBX1 and myocardial markers of differentiation were examined in transfected P19 cells; uc40-siRNA-44 downregulated myocardial markers and upregulated PBX1 expression. These results suggest that uc.40 may play an important role during the differentiation of P19 cells by regulation of PBX1 to promote proliferation and inhibit apoptosis. These studies provide a foundation for further study of uc.40/PBX1 in cardiac development.

The expression profile analysis and functional prediction of lncRNAs in peripheral blood mononuclear cell in maintenance hemodialysis patients developing heart failure

“…Moreover, based on GO and KEGG pathway analyses, we found that target genes of highly differentially expressed lncRNAs were mainly involved in pathways including regulation of actin cytoskeleton, actin dynamics signaling pathway, and so on. Among the 9 target genes of hsa-miR-4505, CRAT [26,27], ITGB5 [19,28,29], SMIM5 [30], CHI3L1 [31], GPC6 [32,33] and PBX1 [34] have been con rmed as the key target genes for cardiac function damage in hemodialysis patients. In addition, functional prediction of the target genes revealed that ITGB5 was enriched in actin dynamics signaling pathway, which was consistent with the results of functional enrichment analysis of related lncRNAs.…”

Research progress of PBX1 in developmental and regenerative medicine

“…Knockdown of uc40-siRNA44 increases the number of cardiac stem cells during P19 cell differentiation. It promotes the proliferation of P19 cells and inhibits apoptosis induced by serum starvation, which may play a role in regulating PBX1 60 .…”

Ultra-conserved RNA: a novel biological tool with diagnostic and therapeutic potential

“…In this mechanism, Mef2c is negatively regulated by uc.167, however, overexpression of Mef2c can reverse the effects of uc.167 [ 10 ]. Uc.40 also has a known role in inhibiting the proliferation and differentiation of cardiomyocytes by downregulating Pre-B-cell leukemia homeobox transcription factor 1 (PBX1) expression [ 11 ]. Interestingly, although uc.245 has no direct effects on the cell cycle, it can play the same role as the ucRNAs discussed above by downregulating the expression of friend of GATA 2 (FOG2).…”