三唑磷对斑马鱼HULC基因的表达调控

Effect of Triazophos on the Expression of HULC Gene in Zebrafish

HULC(highly upregulated in liver cancer)属lncRNA(长链非编码RNA),调控细胞自噬,在肝癌组织中高度上调表达。本文利用生物信息学数据库和实时荧光定量PCR技术对三唑磷处理后斑马鱼中lncRNA HULC调控机制进行研究,进一步了解lncRNA-miRNA-mRNA的调控网络,并且深入了解农药三唑磷对斑马鱼的环境毒理影响。基于芯片技术检测HTT、HOTTIP、NEAT1、NPPA和HULC的表达情况,三唑磷处理斑马鱼前后HTT表达无差异,HOTTIP表达略微下调,NEAT1、NPPA和HULC出现明显上调表达。软件RegRNA 2. 0预测出dre-miR-2185*等7条miRNA与HULC发生相互作用,进一步预测其调控靶基因,构建调控网络。荧光定量PCR结果表明,HULC表达上调1. 5倍、dre-miR-2185*表达下调6. 63倍和离子型谷氨酸受体基因gria1a表达下调2倍。同时三者都可作为生物标志物来监测化学物质对环境和生物体的影响,以期对肝癌等相关疾病提供新的治疗方法。

HULC(highly upregulated in liver cancer), a lncRNA(long chain non-coding RNA), which regulates autophagy, is highly up-regulated in tissue of liver cancer. To further understand the regulation network of lncRNA-miRNA-mRNA, the transcriptional regulatory manner of lncRNA HULC in zebrafish treated with triazophos was studied by using bioinformatics database and real-time fluorescence quantitative PCR(qRT-PCR) in this paper. And give an insight into the effect of pesticide triazophos on zebrafish in environmental toxicology. According to expression of HTT, HOTTIP, NEAT1, NPPA and HULC, which was detected with microarray, no significant difference on the expression of HTT after zebrafish were treated with triazophos, and HOTTIP was slightly down-regulated. NEAT1, NPPA and HULC were significantly up-regulated. Differential expression of 7 miRNAs, including dre-miR-2185*, were predicted with RegRNA 2.0.And we predicted target genes to build a regulatory network. The results of qRT-PCR showed that HULC expression was up-regulated for 1.5 times and dre-miR-2185 * was down-regulated for 6 times, while the expression of ionase glutamate receptor gene gria1a was down-regulated for 2 times. Meanwhile, those three RNAs can be biomarkers to monitor the impact of chemical substances on the environment and organisms, which can be expected to provide new treatments on liver cancer and other related diseases.