Ministry of Education Key Laboratory of Child Development and Disorders, Pediatric Research Institute, Children's Hospital of Chongqing Medical University, Chongqing 400014, P.R. China.
Cardiovascular Department (Internal Medicine), Children's Hospital of Chongqing Medical University, Chongqing 400014, P.R. China.
Int J Mol Med. 2014 May;33(5):1075-82. doi: 10.3892/ijmm.2014.1687. Epub 2014 Mar 6.
The aim of the present study was to investigate the effects of Islet-1 on the process of mesenchymal stem cell (MSC) differentiation into cardiomyocyte-like cells and to elucidate the possible mechanisms involved. Lentiviral vectors expressing Islet-1 (Lenti-Islet-1) were constructed and used for C3H10T1/2 cell transfection. Cell morphology was observed. Cardiac-related genes and proteins were detected by qPCR and western blot analysis. Epigallocatechin gallate (EGCG) was used as an inhibitor of acetylated histone H3 (AcH3). AcH3 was detected by chromatin immunoprecipitation. Cells overexpressing Islet-1 tended to change into fibroblast-like cells and were arranged in the same direction. The enhanced expression of GATA binding protein 4 (Gata4), NK2 homeobox 5 (Nkx2.5), myocyte enhancer factor 2C (Mef2c) and cardiac troponin T (cTnT) was observed in the cells overexpressing Islet-1 following transfection with Lenti-Islet-1. However, the expression of hepatocyte-, bone- and neuronal-specific markers was not affected by Islet-1. The AcH3 relative amount increased following transfection with Lenti-Islet-1, which was associated with the enhanced expression of Gata4, Nkx2.5 and Mef2c in these cells. The expression of Gata4, Nkx2.5 and Mef2c in the C3H10T1/2 cells transfected with Lenti-Islet-1 and treated with EGCG was reduced following treatment with EGCG. The data presented in this study indicate that Islet-1 specifically induces the differentiation of C3H10T1/2 cells into cardiomyocyte-like cells, and one of the mechanisms involved is the regulation of histone acetylation.
本研究旨在探讨胰岛细胞 1（Islet-1）在间充质干细胞（MSC）向心肌样细胞分化过程中的作用，并阐明其可能的作用机制。构建了表达 Islet-1 的慢病毒载体（Lenti-Islet-1），并用于 C3H10T1/2 细胞转染。观察细胞形态。通过 qPCR 和 Western blot 分析检测心脏相关基因和蛋白。表没食子儿茶素没食子酸酯（EGCG）被用作乙酰化组蛋白 H3（AcH3）的抑制剂。通过染色质免疫沉淀检测 AcH3。过表达 Islet-1 的细胞倾向于变成成纤维细胞样细胞，并排列在同一方向。转染 Lenti-Islet-1 后，过表达 Islet-1 的细胞中 GATA 结合蛋白 4（Gata4）、NK2 同源盒 5（Nkx2.5）、肌细胞增强因子 2C（Mef2c）和心肌肌钙蛋白 T（cTnT）的表达增强。然而，Islet-1 对肝细胞、骨细胞和神经元特异性标志物的表达没有影响。转染 Lenti-Islet-1 后，AcH3 相对量增加，与这些细胞中 Gata4、Nkx2.5 和 Mef2c 的表达增强有关。用 EGCG 处理转染了 Lenti-Islet-1 并经 EGCG 处理的 C3H10T1/2 细胞中 Gata4、Nkx2.5 和 Mef2c 的表达降低。本研究的数据表明，Islet-1 特异性诱导 C3H10T1/2 细胞向心肌样细胞分化，其机制之一是调节组蛋白乙酰化。
