三唑磷对斑马鱼H19和Sox2OT基因表达调控的影响

Effect of triazophos on the expression of H19 and Sox2OT genes in Danio rerio

长链非编码RNA (long non-coding RNA, lncRNA)是指长度大于200 nt且不具有编码蛋白质功能的一类RNA分子。当生物体暴露于环境污染物时,其lncRNA通常会异常表达,并与癌症等疾病的发生相关。采用亚致死浓度的三唑磷处理斑马鱼后,利用人源基因芯片初步筛选出了异常表达的斑马鱼lncRNA,并通过生物信息学软件预测了与lncRNA相互作用的微RNA(micro RNA, miRNA)及其靶基因,最后通过实时荧光定量PCR验证了该miRNA及其靶基因在斑马鱼中的表达,从而构建起了长链非编码RNA-微RNA-信使RNA (lncRNA-miRNA-mRNA)的调控网络。荧光定量PCR检测结果表明:与空白对照组相比,经三唑磷处理后,斑马鱼lncRNA Sox2OT基因的表达下调了35%;lncRNA H19基因的表达上调了1.41倍,而与其可能存在相互作用的dre-let-7c基因的表达则下调了78%,同时dre-let-7c的靶基因ddx18的表达上调了2.11倍;差异均达显著水平(P <0.05)。据此推测,在三唑磷的作用下,lncRNA H19可能作为靶基因ddx18的竞争性内源RNA而与dre-let-7c发生相互作用,从而调控靶基因ddx18的表达。因此,lncRNA H19、dre-let-7c及ddx18都有可能作为生物标志物用于监测三唑磷对环境和生物体的影响。

Long non-coding RNAs(lncRNAs)are defined as a class of RNA molecule with a length of over 200 nucleotides and without protein-coding function.On the exposure to environmental pollutants,dysregulated expression of lncRNAs has been observed,which is associated with several diseases such as cancer.A human-derived microarray was used to screen the zebrafish(Danio rerio)lncRNA of zebrafish after sublethal triazophos treatment.The miRNAs and their target genes were predicted by the bioinformatics software.Finally,real-time fluorescent quantitative PCR was used to verify their relative expression in zebrafish,so that a lncRNA-miRNA-mRNA regulatory network was constructed.Results of real-time PCR showed that lncRNA Sox2 OT was reduced by 35%after triazophos treatment compared with that of the control group.The expression of lncRNA H19 was up-regulated by 1.41-fold,while that of dre-let-7 c was reduced by 78%.Furthermore,the expression of ddx18,which may be the target gene of dre-let-7 c,was up-regulated by 2.11-fold.The difference was significant(P<0.05).It’s proposed that lncRNA H19 may interact with dre-let-7 c as a competitive endogenous RNA to regulate the expression of ddx18 after the triazophos treatment.Therefore,lncRNA H19,dre-let-7 c and ddx18 can be potentially used as biomarkers to monitor the effects of triazophos on the environment and organisms.