Impact of miR-26b on cardiomyocyte differentiation in P19 cells through regulating canonical/non-canonical Wnt signalling
Impact of miR-26b on cardiomyocyte differentiation in P19 cells through regulating canonical/non-canonical Wnt signalling
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miR-26b通过调节经典/非经典Wnt信号对P19细胞心肌细胞分化的影响
DOI：
10.1111/cpr.12371
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发表时间：
2017
影响因子：
8.5
通讯作者：
Peng Luying
中科院分区：
文献类型：
--
作者：
Wang Duo;Liu Chang;Wang Yumei;Wang Wenjing;Wang Kang;Wu Xiujuan;Li Zhigang;Zhao Cuimei;Li Li;Peng Luying
背景与目的心肌细胞分化的调控与microRNAs（miRNAs）密切相关，miRNAs在心脏发育过程中发挥着重要作用。然而，miRNAs在早期心脏发育中介导的调控机制仍然是推测性的。在这里，我们评估了miR-26 b在P19细胞系心肌细胞分化过程中的影响。材料和方法通过慢病毒载体转导，在P19细胞中过表达miR-26 b。采用定量真实的时间PCR检测P19细胞分化过程中心脏相关基因的mRNA丰度，采用Western印迹检测蛋白表达。ICG-001用于阐明β-catenin对P19细胞分化的作用。使用细胞计数试剂盒-8（CCK-8）监测细胞增殖。结果miR-26 b的过表达通过影响Wnt 5a信号和Gsk 3 β活性，上调Gata 4、cTNT、α-MHC和α-Actinin等心肌细胞相关基因的表达水平，全面反映心肌细胞分化。然而，ICG-001沿着细胞增殖的抑制而阻断分化。此外，miR-26 b还通过调控CyclinD 1促进P19细胞增殖，从而证明P19细胞快速聚集并向心肌样细胞分化.ConclusionsOur结果表明，miR-26 b通过调控经典和非经典Wnt信号通路促进P19细胞向心肌样细胞分化。
Background and objectivesThe control of cardiomyocyte differentiation is tightly linked to microRNAs (miRNAs), which have been emerging as important players in heart development. However, the regulation mechanisms mediated by miRNAs in early heart development remains speculative. Here, we evaluated the impact of miR‐26b during the progression of cardiomyocyte differentiation from the P19 cell line.Materials and methodsThe overexpression of miR‐26b in P19 cells was performed by transduction with lentivirus vector. The levels of cardiac‐related genes during P19 cell differentiation were detected using quantitative real‐time PCR for mRNA abundance and Western blots for protein expression. ICG‐001 was applied to elucidate the role of β‐catenin on P19 cells differentiation. The Cell Counting kit‐8 (CCK‐8) was used to monitor the cell proliferation. The target genes of miR‐26b were validated using the dual luciferase reporter system.ResultsOverexpression of miR‐26b upregulates the expression level of cardiomyocyte‐related genes such as Gata4, cTNT, α‐MHC and α‐Actinin that comprehensively represent cardiomyocyte differentiation by effecting Wnt5a signalling and Gsk3β activity. However, ICG‐001 blocks the differentiation along with inhibition of the cell proliferation. In addition, miR‐26b also regulates CyclinD1 to promote P19 cell proliferation, thereby, demonstrating the rapid aggregation and differentiation programming of these cells into cardiomyocytic types.ConclusionsOur results indicated that miR‐26b exerts a role on promoting cardiomyocyte differentiation of P19 cells by controlling the canonical and non‐canonical Wnt signalling.