Effects of long non-coding RNA uc.245 on cardiomyocyte-like differentiation in P19 cells via FOG2
Effects of long non-coding RNA uc.245 on cardiomyocyte-like differentiation in P19 cells via FOG2
复制标题
长非编码RNA uc.245通过FOG2对P19细胞心肌细胞样分化的影响
DOI：
10.1016/j.gene.2018.12.080
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发表时间：
2019-04-30
期刊：
影响因子：
3.5
通讯作者：
Li, Mengmeng
中科院分区：
文献类型：
--
作者：
Liu, Heng;Hu, Yin;Li, Mengmeng
每年，心脏病都可能在全世界范围内导致高发病率和死亡率。含有超保守元件（UCE）的长非编码RNA（IncRNA）可能在心肌细胞分化中发挥重要作用。有必要进一步研究IncRNA-UC调节胚胎心脏发育的潜在机制。在本研究中，我们研究了Inc-uc.245对DMSO刺激下P19细胞增殖、迁移、凋亡和心肌细胞样分化的影响，并假设inc-uc.245会通过FOG2影响心肌细胞分化。将pGPU6/GFP/Neo-uc.245和pGPU6/GFP/Neo-shRNA-uc.245慢病毒载体分别转染P19细胞，过表达或沉默uc.245。 MIT实验、Annexin V-FITC/PI双染、划痕实验和Transwell实验结果表明，uc.245过表达可显着抑制P19细胞增殖、迁移、心肌细胞样分化，但促进细胞凋亡。相反，sh-uc.245处理引起相反的变化。 Uc.245过表达明显下调心肌特异性分子标志物（cTnI、ANP、α-MHC、Nkx2.5、GATA4、MEF2C）的表达，但显着上调FOG2的表达。随后，我们将装载FOG2或shRRNA-FOG2的重组载体转染至P19细胞中，以进一步探讨FOG2在uc.245调节的心肌细胞样分化中的功能意义。有趣的是，我们发现在uc.245过表达和沉默的P19细胞中，FOG2的过表达促进细胞增殖、迁移并抑制细胞凋亡，特别是在uc.245沉默的细胞系中。此外，在 uc.245 过表达和沉默的 P19 细胞中，sh-FOG2 促进心肌细胞样分化，并在基因和蛋白质水平上调心肌生成特异性标记物的表达。同样，sh-FOG2 的这种上调效应在 uc.245 沉默后更加明显。这些发现表明，FOG2 是 uc.245 调节的 P19 细胞分化为心肌细胞过程中的关键介质。预计lnc-uc.245/FOG2将成为心脏病的有前景的治疗靶点。
Each year, cardiac diseases may cause a high morbidity and mortality worldwide. Long non-coding RNAs (IncRNAs) that contained ultra-conserved elements (UCEs) may play important roles on cardiomyocytes differentiation. Further investigations underlying mechanisms of IncRNA-UC regulating embryonic heart development are necessary.In this study, we investigated the effects of Inc-uc.245 on proliferation, migration, apoptosis, and cardiomyocyte-like differentiation in P19 cells with DMSO stimulation, and hypothesized that inc-uc.245 would influence cardiomyocytes differentiation via FOG2. Lentiviral vectors of pGPU6/GFP/Neo-uc.245 and pGPU6/GFP/Neo-shRNA-uc.245 were respectively transfected into P19 cells to overexpress or silence uc.245. MIT assay, Annexin V-FITC/PI double-staining, scratch test and transwell assay were performed and the results showed that uc.245 overexpression could significantly suppress P19 cell proliferation, migration, cardiomyocyte-like differentiation but promote cell apoptosis. Contrarily, sh-uc.245 treatment caused the opposite changes. Uc.245 overexpression obviously downregulated the expression of cardiomyogenic-specific molecular markers (cTnI, ANP, alpha-MHC, Nkx2.5, GATA4, MEF2C) but remarkably upregulated the expression of FOG2. Subsequently, we transfected the recombinant vectors loaded FOG2 or shRRNA-FOG2 into P19 cells to further address the functional significance of FOG2 in uc.245-regulated cardiomyocyte-like differentiation. Interestingly, we found that overexpressing of FOG2 promoted cell proliferation, migration, and inhibited apoptosis both in uc.245 overexpressed and silenced P19 cells, especially in uc.245 silenced cell line. In addition, sh-FOG2 promoted cardiomyocyte-like differentiation and upregulated the expression of cardiomyogenic-specific markers at the gene and protein levels both in uc.245 overexpressed and silenced P19 cells. Similarly, this upregulation effect of sh-FOG2 was more obvious after uc.245 silencing.These findings suggest that FOG2 is a key mediator during uc.245-regulated differentiation of P19 cells into cardiomyocytes. It is expected that lnc-uc.245/FOG2 will become a promising therapeutic target for cardiac diseases.