六价铬职业接触人群血浆DNA损伤反应相关microRNA的鉴定及功能研究

项目名称： 六价铬职业接触人群血浆DNA损伤反应相关microRNA的鉴定及功能研究

项目编号： No.81472960

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 楼建林

作者单位： 浙江省医学科学院

项目金额： 70万元

中文摘要： 职业接触六价铬可通过诱发DNA损伤致多种健康损害，但机制仍不清楚。研究显示，microRNA在调节DNA损伤反应中起重要作用。我们前期研究发现，铬盐职业接触工人血浆miR-340、miR-151和miR590的表达上调，提示miRNA参与六价铬诱导的DNA损伤反应。因此，本项目拟在铬盐职业接触人群中，用QuantiGene核酸定量技术检出DNA损伤反应相关差异基因；用芯片技术在小样本人群中筛出差异microRNAs，经关联分析后确定DNA损伤反应相关microRNAs，并用定量PCR方法在大样本人群中验证；在体外试验中，逆转表达后研究microRNAs在调控六价铬诱发DNA损伤反应中的作用，并用双荧光素酶报告基因法和回复试验验证靶基因。项目预期确定可作为铬盐职业接触人群生物标志物的microRNAs及靶基因，明确其在DNA损伤反应中的作用及机制，为六价铬毒作用的机制研究和预防提供新的思路。

中文关键词： 职业流行病学；DNA损伤反应；微小RNA

英文摘要： Occupational exposure to hexavalent chromium (Cr (Ⅵ)) leads to various kinds of health damage by inducing DNA damage, but the underlying mechanims are not clear yet. Recent studies found that microRNA played an important role in regulating DNA damage response. Our previous study suggested that the expression levels of Plasma miR-340, miR-151, and miR-590 were up-regulated in workers occupationally exposed to Cr (Ⅵ), which indicated that microRNA might take part in DNA damage response induced by Cr (Ⅵ). Therefore, in the present study, single strand DNA breaks and double strand DNA breaks were studied in workers occupationally exposed to chromate and controls, and QuantiGene 80 Plex Assay was used to found differentially expressed genes related to DNA damage response; microRNA microarray is used to screen differential microRNAs in small sample size populations, and differentially expressed microRNAs related to DNA damage response will be confirmed after correlation analysis between differential genes and differential microRNAs, and the expression of differential miRNAs is validated in large sample size populations; The function of microRNAs is studied once their expression difference are confirmed, and the role of miRNAs in regulating DNA damage response induced by Cr (Ⅵ) is studied by reversing the expression of miRNAs, and the expression of target genes is validated by Dual Luciferase Reporter Gene Assay and rescuing assay. The aim of our study is to discover microRNAs and their target genes as biomarkers for workers occupationally exposed to Cr (Ⅵ), and to study the role of these microRNAs in response to DNA damage, providing a new road to mechanim study and prevention of the toxicity of Cr (Ⅵ).

英文关键词： Occupational epidemiology;DNA damage response;MicroRNA