Serum DNase I activity can be used as a sensitive marker for detection of acute myocardial infarction

血清DNase I活性可作为检测急性心肌梗死的敏感标志物

基本信息

批准号：
16209023
负责人：
KOMINATO Yoshihiko
金额：
$ 28.45万
依托单位：
Gunma University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16209023/
关键词：
Deoxyribonuclease I QGP-1 cell Hypoxia Transcription Myocardial infarction Promoter 膵臓癌細胞GSP1

项目摘要

We demonstrated that serum DNase I activity could be used as novel diagnostic marker for the early detection of acute myocardial infarction (AMI) ; abrupt elevation of serum DNase I activity levels occurs within 3h of the onset of symptoms in patients with AMI, permitting the diagnosis of AMI before accurate CK-MB and c-TnT results become available. Moreover, we investigated alterations in serum DNase I levels after transient ischemia induced during percutaneous coronary intervention(PCI). Serum DNase I activity had risen significantly from base line by 3h after completion of the PCI procedure. However, the mechanism for the elevation of serum DNase I activity induced by ischemia during AMI or PCI remain to be elucidated. To elucidate the molecular basis of this phenomenon, it is important to understand the regulatory mechanism of the human DNase I gene(DNASE1) expression. We first mapped the transcription start sites of DNASE1 in human pancreas and in the DNase I producing human pancr … More eatic cancer cell line QGP-1, and revealed a novel site 12kb upstream of exon 1, which was previously believed to be the single transcription starting exon. This initiation site markes an alternative starting exon, designated 1a. Exon 1 and 1a were used simultaneously as transcription starting exon in pancreas and QGP-1 cells. Promoter assay EMSA and chromatin immunoprecipitation analysis with QGP-1 cell lines showed that the promoter region of exon 1a in which the Sp1 transcription factor is specifically involved in promoter activity. This is the first to be identified as a transcription factor responsible for gene expression of vertebrate DNase I gene. Furthermore, RT-PCR analysis indicated alternative splicing of human DNase 1 pre-mRNA in pancreas and QGP-1 cells. Only two transcripts among eight alternative splicing products identified can be translated to produce intact DNase I protein. These results suggested that human DNase 1 expression is regulated through the use of alternative promoter and alternative splicing. Less

我们证明，血清 DNase I 活性可以作为早期检测急性心肌梗死 (AMI) 的新诊断标志物；AMI 患者出现症状后 3 小时内血清 DNase I 活性水平突然升高，从而可以进行诊断。在获得准确的 CK-MB 和 c-TnT 结果之前，我们研究了 AMI 的情况。此外，我们还研究了经皮冠状动脉介入治疗 (PCI) 期间诱导的短暂性缺血后血清 DNase I 水平的变化。 PCI 手术完成后 3 小时，血清 DNase I 活性较基线显着升高，但 AMI 或 PCI 期间缺血引起的血清 DNase I 活性升高的机制仍有待阐明。了解人类 DNase I 基因 (DNASE1) 表达的调控机制非常重要。我们首先绘制了人类胰腺和产生人类胰腺中 DNASE1 的转录起始位点。食癌细胞系 QGP-1，并揭示了外显子 1 上游 12kb 的新位点，该位点先前被认为是单个转录起始外显子，该起始位点标记了同时使用的替代起始外显子，称为 1a 和 1a。作为胰腺和 QGP-1 细胞中转录起始外显子的启动子测定对 QGP-1 细胞系的 EMSA 和染色质免疫沉淀分析表明，外显子的启动子区域。 1a，其中 Sp1 转录因子专门参与启动子活性，这是第一个被鉴定为负责脊椎动物 DNase I 基因表达的转录因子。此外，RT-PCR 分析表明人类 DNase 1 前体存在选择性剪接。胰腺和 QGP-1 细胞中的 mRNA。在八种选择性剪接产物中，只有两种转录物可以翻译产生完整的 DNase I 蛋白。这些结果表明，人类 DNase 1 的表达是通过使用来调节的。选择性启动子和选择性剪接较少。