三唑磷对斑马鱼miR-99及其预测靶标ckmt2的调控作用的研究

项目名称： 三唑磷对斑马鱼miR-99及其预测靶标ckmt2的调控作用的研究

项目编号： No.21307113

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 环境科学、安全科学

项目作者： 张凯

作者单位： 浙江理工大学

项目金额： 25万元

中文摘要： 肌酸激酶(Creatine kinase,CK)能够可逆的催化肌酸与ATP之间的转磷酰基反应。近年来发现，肌酸激酶可作为肿瘤诊断的标志酶。MicroRNA是一类长度约为20-23nt的非编码小RNA，在转录后水平对基因表达水平进行调控，是癌症发生、发展及化学物质毒理学等过程中重要的表观遗传调控因子。我们的研究结果表明，三唑磷可引起斑马鱼体内miR-99及其潜在靶基因ckmt2的表达发生变化。这是否预示三唑磷具有潜在的致癌效应，尚未见相关报道。本项目拟以斑马鱼miR-99为对象，采用实时荧光定量 PCR、细胞转染等技术，研究三唑磷处理后斑马鱼miR-99与与ckmt2的共表达变化、miR-99与ckmt2基因3'-UTR间的相互作用和miR-99对CKmt2蛋白表达的调控作用，旨在阐明miR-99和ckmt2在异源化学物质毒理学效应中的作用与关系。

中文关键词： ckmt2；斑马鱼；miR-99；miR-217；毒理学效应

英文摘要： Creatine kinase (CK) is an enzyme that catalyzes the reversible conversion of ADP and phosphocreatine into ATP and creatine, and was reported to be tumor marker in the diagnosis of some primary cancer. MicroRNAs (miRNAs) are a class of small (20-23 nucleotides), endogenous and non-coding RNAs, emerged as key post-transcriptional regulators of gene expression, and directly involved in cancer occurrence, development, and process of chemical toxicology. Our latest results shown the expression of miR-99 and its predicted target gene ckmt2 in zebrafish will change after treated with triazophos. This is the only report shown triazophos has potential carcinogenicity. Numerous studies must be conducted to determine the hazards of the particular pesticide and its use.We propose to investigate the co-expression alteration of miR-99 and ckmt2, and the interaction of miR-99 and 3'-UTR of ckmt2 after treated with triazophos in zebrafish. Try to figure out the mechanism of the regulation ckmt2 by miR-99 with quantitative RT-PCR and cell transfection techniques. With this study, we can clarify the interaction of miR-99 and ckmt2 in the toxicology effects of heterologous chemicals and we can also classify the carcinogenic potential of the pesticides triazophos.

英文关键词： ckmt2；zebrafish；miR-99；miR-217；toxicological effect