Yu Zhou, Shuang Gao, Liangcai Ding, Han Yan, Shuchao Pang, Bo Yan
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引用次数: 0
Abstract
Dilated cardiomyopathy (DCM) is caused by a combination of genetic susceptibility and environmental factors. Cathepsin B affects the pathogenesis of DCM; however, its molecular mechanism is still unclear. In this study, we examined the association of rare CTSB variants with the occurrence of DCM. This case-control study involved 394 participants: 142 patients with DCM and 252 healthy controls. DNA was extracted from the peripheral leukocytes of all participants, and CTSB variants were analyzed and identified using polymerase chain reaction amplification. Functional analysis was performed using the dual-luciferase reporter assay, and the ability of genetic CTSB variants to bind to transcription factors (TFs) was analyzed and validated using the electrophoretic mobility shift assay (EMSA). Two single-nucleotide polymorphisms (SNPs) were identified in the study population. One SNP, g.4803 T > C (rs1293312), was more common in patients with DCM. A second SNP, g.4954 T > A (rs942670850), was identified in two patients with DCM. Both SNPs significantly enhanced the transcriptional activity of CTSB promoters. An analysis using the TRANSFAC database revealed that these SNPs affect TF binding, which was confirmed using the EMSA. Our results demonstrate that within the CTSB promoter, the genetic variants g.4803T>C (rs1293312) and g.4954 T > A (rs942670850) are rare risk factors for DCM development.
扩张型心肌病(DCM)由遗传易感性和环境因素共同引起。组织蛋白酶B影响DCM的发病机制;然而,其分子机制尚不清楚。在这项研究中,我们研究了罕见的CTSB变异与DCM发生的关系。这项病例对照研究涉及394名参与者:142名DCM患者和252名健康对照者。从所有参与者的外周白细胞中提取DNA,并使用聚合酶链反应扩增分析和鉴定CTSB变异。使用双荧光素酶报告基因法进行功能分析,并使用电泳迁移迁移试验(EMSA)分析和验证遗传CTSB变异与转录因子(TFs)结合的能力。在研究人群中发现了两个单核苷酸多态性(snp)。一个SNP, g.4803T > C (rs1293312)在DCM患者中更为常见。第二个SNP, g.4954在2例DCM患者中鉴定出T > A (rs942670850)。这两个snp都显著增强了CTSB启动子的转录活性。使用TRANSFAC数据库进行的分析显示,这些snp影响TF结合,这一点通过EMSA得到了证实。我们的研究结果表明,在CTSB启动子内,遗传变异g.4803T>C (rs1293312)和g.4954T > A (rs942670850)是DCM发展的罕见危险因素。
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