Ge Yinghua, Zhang Jiewen, Shi Xinyi, Lu Conger, Yang Lingrong, Li Yuanyuan, Chen Yanna, Cheng Dandan, Bai Jing, Lv Zhengbing, Liu Lili
College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou, 310018, China.
Zhejiang Provincial Key Laboratory of Silkworm Bioreactor and Biomedicine, Hangzhou, 310018, China.
Fish Physiol Biochem. 2017 Jun;43(3):791-802. doi: 10.1007/s10695-016-0332-1. Epub 2017 Jan 4.
Liver regeneration is a complicated process, and understanding the regulatory mechanism will be helpful in the treatment of diseases associated with liver. In this study, the one-third liver resection model was established in Chiloscyllium plagiosum, and the whole transcriptome of the C. plagiosum was generated using the Illumina-Solexa sequencing platform. Differentially expressed genes were analyzed using bioinformatics methods and verified using quantitative real-time PCR (qRT-PCR). Using miRanda and TargetScan, we screened the microRNA library for miRNAs that target the glutathione S-transferase P1(GSTP1) gene. Dual-luciferase reporter assays were used to confirm binding between the miRNA and GSTP1. Finally, we used western blotting analysis to determine expression of the GSTP1 protein. As a result, 65,356 unigenes were obtained in normal and damaged liver tissues, with mean length of 955 bp. A total of 359 differentially expressed genes were acquired; 217 of which were upregulated, and 142 were downregulated, including the GSTP1 gene, following liver resection. The presence of the GSTP1 protein in C. plagiosum was shown for the first time. Luciferase reporter assay revealed that GSTP1 messenger RNA was targeted by ipu-miR-143. The discovery and differential expression analysis of GSTP1 in C. plagiosum will be a valuable resource to explain the molecular mechanism of GSTP1 regulation of liver repair.
肝脏再生是一个复杂的过程，了解其调控机制将有助于治疗与肝脏相关的疾病。在本研究中，建立了斜带石斑鱼三分之一肝切除模型，并使用Illumina-Solexa测序平台生成了斜带石斑鱼的全转录组。利用生物信息学方法分析差异表达基因，并通过定量实时PCR（qRT-PCR）进行验证。使用miRanda和TargetScan，我们在microRNA文库中筛选了靶向谷胱甘肽S-转移酶P1（GSTP1）基因的miRNA。采用双荧光素酶报告基因检测法确认miRNA与GSTP1之间的结合。最后，我们使用蛋白质免疫印迹分析来确定GSTP1蛋白的表达。结果，在正常和受损肝脏组织中获得了65356个单基因，平均长度为955bp。共获得359个差异表达基因；其中217个上调，142个下调，包括肝切除后的GSTP1基因。首次在斜带石斑鱼中显示了GSTP1蛋白的存在。荧光素酶报告基因检测显示GSTP1信使核糖核酸被ipu-miR-143靶向。斜带石斑鱼中GSTP1的发现和差异表达分析将为解释GSTP1调控肝脏修复的分子机制提供有价值的资源。
