Department of Cardiovascular Medicine, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan.
J Cell Biochem. 2012 Nov;113(11):3455-65. doi: 10.1002/jcb.24222.
Tissue-specific patterns of gene expression play an important role in the distinctive features of each organ. Small CTD phosphatases (SCPs) 1-3 are recruited by repressor element 1 (RE-1)-silencing transcription factor/neuron-restrictive silencer factor (REST/NRSF) to neuronal genes that contain RE-1 elements, leading to neuronal gene silencing in non-neuronal cells. SCPs are highly expressed in the heart and contain microRNAs (miR)-26b, 26a-2, and 26a-1 with the same seed sequence in their introns. Therefore, we tried to investigate the roles of miR-26b and its host gene in neonatal rat cardiomyocytes. Overexpression of miR-26b suppressed the mRNA expression levels of ANF, βMHC, and ACTA1 and reduced the cell surface area in cardiomyocytes. We confirmed that miR-26b targets the 3' untranslated region (3'UTR) of GATA4 and canonical transient receptor potential channel (TRPC) 3. Conversely, silencing of the endogenous miR-26b family enhanced the expression levels of TRPC3 and GATA4. On the other hand, overexpression of SCP1 induced the mRNA expression of ANF and βMHC and increased the cell surface area in cardiomyocytes. Next, we compared the effect of overexpression of SCP1 with its introns and SCP1 cDNA to observe the net function of SCP1 expression on cardiac hypertrophy. When the expression levels of SCP1 were the same, the overexpression of SCP1 cDNA had a greater effect at inducing cardiac hypertrophy than SCP1 cDNA with its intron. In conclusion, SCP1 itself has the potential to induce cardiac hypertrophy; however, the effect is suppressed by intronic miR-26b in cardiomyocytes. miR-26b has an antagonistic effect on its host gene SCP1.
组织特异性基因表达模式在每个器官的独特特征中起着重要作用。小 CTD 磷酸酶(SCPs)1-3 通过抑制元件 1(RE-1)-沉默转录因子/神经元限制沉默因子(REST/NRSF)招募到含有 RE-1 元件的神经元基因,导致非神经元细胞中的神经元基因沉默。SCPs 在心脏中高度表达,并且在其内含子中含有相同种子序列的 microRNAs(miR)-26b、26a-2 和 26a-1。因此,我们试图研究 miR-26b 及其宿主基因在新生大鼠心肌细胞中的作用。miR-26b 的过表达抑制了心肌细胞中 ANF、βMHC 和 ACTA1 的 mRNA 表达水平,并减小了细胞表面积。我们证实 miR-26b 靶向 GATA4 和经典瞬时受体电位通道(TRPC)3 的 3'非翻译区(3'UTR)。相反,内源性 miR-26b 家族的沉默增强了 TRPC3 和 GATA4 的表达水平。另一方面,SCP1 的过表达诱导了 ANF 和βMHC 的 mRNA 表达,并增加了心肌细胞的表面积。接下来,我们比较了 SCP1 及其内含子和 SCP1 cDNA 的过表达的效果,以观察 SCP1 表达对心肌肥大的净功能。当 SCP1 的表达水平相同时,SCP1 cDNA 的过表达对诱导心肌肥大的作用大于带有内含子的 SCP1 cDNA。总之,SCP1 本身具有诱导心肌肥大的潜力;然而,在心肌细胞中,内含子 miR-26b 抑制了这种作用。miR-26b 对其宿主基因 SCP1 具有拮抗作用。