三唑磷对斑马鱼H19和Sox2OT基因表达调控的影响

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。

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

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 Sox2OT 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-7c was reduced by 78%. Furthermore, the expression of ddx18, which may be the target gene of dre-let-7c, 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-7c as a competitive endogenous RNA to regulate the expression of ddx18 after the triazophos treatment. Therefore, lncRNA H19, dre-let-7c and ddx18 can be potentially used as biomarkers to monitor the effects of triazophos on the environment and organisms.