Coronary artery disease (CAD) has been the leading cause of morbidity and mortality worldwide, and its pathogenesis is closely related with the proliferation and migration of vascular smooth muscle cell (VSMC). We previously reported a truncated GATA4 protein lacking C-terminus induced by p.S335X mutation in cardiomyocyte from ventricular septal defect (VSD) patients. However, it is still unclear whether GATA4 p.S335X mutation could influence the development of CAD. GATA4 wild-type (WT) and p.S335X mutant (MU) overexpression plasmids were constructed and transfected transiently into rat coronary artery smooth muscle cell (RCSMC) to observe the proliferative and migratory abilities by MTS and wound healing assay, respectively. PCR array was used to preliminarily detect the expression of phenotypic modulation-related genes, and QRT-PCR was then carried out to verify the screened differentially expressed genes (DEGs). The results showed that, when stimulated by fetal bovine serum (10%) for 24 h or tumor necrosis factor-α (10 or 30 ng/ml) for 10 or 24 h, deletion of GATA4 C-terminus by p.S335X mutation in GATA4 enhanced the proliferation of RCSMC, without alteration of the migration capability. Twelve DEGs, including Fas, Hbegf, Itga5, Aimp1, Cxcl1, Il15, Il2rg, Il7, Tnfsf10, Il1r1, Irak1, and Tlr3, were screened and identified as phenotypic modulation-related genes. Our data might be beneficial for further exploration regarding the mechanisms of GATA4 p.S335X mutation on the phenotypic modulation of coronary VSMC.

中文翻译：

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p.S335X 突变导致 GATA4 C 末端缺失调节冠状动脉血管平滑肌细胞表型

冠状动脉疾病（CAD）已成为全球发病率和死亡率的首要原因，其发病机制与血管平滑肌细胞（VSMC）的增殖和迁移密切相关。我们之前报道了一种缺乏 C 末端的截断 GATA4 蛋白，这是由室间隔缺损 (VSD) 患者的心肌细胞中的 p.S335X 突变引起的。然而，尚不清楚 GATA4 p.S335X 突变是否会影响 CAD 的发展。构建GATA4野生型（WT）和p.S335X突变体（MU）过表达质粒并瞬时转染大鼠冠状动脉平滑肌细胞（RCSMC），分别通过MTS和伤口愈合试验观察增殖和迁移能力。PCR阵列用于初步检测表型调控相关基因的表达，然后进行QRT-PCR以验证筛选的差异表达基因（DEGs）。结果表明，当胎牛血清（10%）刺激24 h或肿瘤坏死因子-α（10 或 30 ng/ml）持续 10 或 24 小时，通过 GATA4 中的 p.S335X 突变删除 GATA4 C 末端可增强 RCSMC 的增殖，而不会改变迁移能力。筛选并鉴定了 12 个 DEG，包括 Fas、Hbegf、Itga5、Aimp1、Cxcl1、Il15、Il2rg、Il7、Tnfsf10、Il1r1、Irak1 和 Tlr3，并鉴定为表型调节相关基因。我们的数据可能有助于进一步探索 GATA4 p.S335X 突变对冠状动脉 VSMC 表型调节的机制。