獭兔(Oryctolagus cuniculus)SLC7A11基因在黑色素细胞中应答基因的筛选及分析  

Screening and Analysis of Response Genes of SLC7A11 in Melanocytes of Rex Rabbit(Oryctolagus cuniculus)

作　　者：卢婷婷 戴莹莹 刘禹飞 刘永琪 白少成 赵博昊 吴信生[1] 陈阳[1] LU Tingting;DAI Yingying;LIU Yufei;LIU Yongqi;BAI Shaocheng;ZHAO Bohao;WU Xinsheng;CHEN Yang(College of Animal Science and Technology,Yangzhou University,Yangzhou,225009)

机构地区：[1]扬州大学动物科学与技术学院,扬州225009

出　　处：《基因组学与应用生物学》2023年第4期384-392,共9页Genomics and Applied Biology

基　　金：国家自然科学基金项目(32172722);扬州大学大学生科创基金项目(X20220652);国家现代农业产业技术体系项目(CARS-43-A-1)共同资助

摘　　要：溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11)在动物皮肤和毛发的黑色素沉积过程中发挥了重要作用。本研究利用Illumina HiSeq高通量测序技术分析獭兔(Oryctolagus cuniculus)SLC7A11基因在黑色素细胞(melanocytes)中的下游应答基因。在黑色素细胞中过表达SLC7A11基因后,与对照组相比,共筛选到110个差异表达基因(differentially expressed genes)(|log2fold change|>1,P<0.05),其中60个上调,50个下调。GO和KEGG富集分析表明,这些差异表达基因主要富集于铁离子的运输、脂质代谢通路。随机选取6个差异表达基因进行实时定量PCR(real time quantitative PCR)验证,与测序结果一致,证明测序结果的可靠性。进一步分析铁离子运输相关基因SLC40A1、SCARA5,以及脂质代谢相关基因PLA2G3、PLA2G2,实时定量PCR结果表明,黑色素细胞中过表达SLC7A11基因显著下调了SLC40A1、SCARA5、PLA2G3、PLA2G2的表达(P<0.05),同时C11 BODIPY 581/591结果表明,过表达SLC7A11基因能够抑制黑色素细胞脂质过氧化。研究结果为进一步阐明SLC7A11影响黑色素细胞的应答机制提供了线索。Solute carrier family 7 member 11(SLC7A11),plays an essential role in the deposition of melanin in animal skin and hair.Using Illumina HiSeq sequencing,we identified the downstream genes and signaling pathways respond to Rex rabbit(Oryctolagus cuniculus)SLC7A11 in melanocytes.A total of 110 differentially expressed genes(|log2fold change|>1,P<0.05)were identified in melanocytes of overexpression SLC7A11 compared to controls,in which 60 were up-regulated and 50 were down-regulated.GO and KEGG analyses revealed that differentially expressed genes were mainly enriched in iron ion transmembrane transport and lipid metabolism pathways.The expression levels of six differentially expressed genes were validated by real time quantitative PCR,and the trends were consistent with the sequencing results,demonstrating the reliability of the sequencing results.The iron transport-related genes SLC40A1,SCARA5,and lipid metabolism-related genesPLA2G3 andPLA2G2 were further selected.Real time quantitative PCR results showed that overexpression of theSLC7A11 gene significantly down-reguted the expression ofSLC40A1,SCARA5,PLA2G3,and PLA2G2(P<0.05),while C11 BODIPY 581/591 results indicated that overexpression ofSLC7A11 gene inhibited lipid peroxidation in melanocytes.The findings might help researchers better understand how SLC7A11 affects melanocytes.

关 键 词：SLC7A11 黑色素细胞 RNA测序 应答基因 

分 类 号：S829.1[农业科学—畜牧学]