LncRNA TCONS_00153149基因在酉州乌羊皮肤黑色素沉积过程中的作用  

Effect of LncRNA TCONS_00153149 Gene in the Process of Melanin Deposition in Youzhou Black Goat(Capra hircus)

作　　者：张丽[1] 李杰[1] 付琳 刘丽 杨恒[3] 王高富[1] 任航行[1] 周鹏[1] 董贤文 ZHANG Li;LI Jie;FU Lin;LIU Li;YANG Heng;WANG Gao-Fu;REN Hang-Xing;ZHOU Peng;DONG Xian-Wen(Chongqing Academy of Animal Sciences/Chongqing Engineering Research Center for Goat,Rongchang 402460,China;Chongqing Chemical Industry Vocational College,Changshou 401228,China;College of Veterinary Medicine,Southwest University,Rongchang 402460,China)

机构地区：[1]重庆市畜牧科学院/重庆市山羊工程技术研究中心,荣昌402460 [2]重庆化工职业学院,长寿401228 [3]西南大学动物医学院,荣昌402460

出　　处：《农业生物技术学报》2022年第5期908-917,共10页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(32002206);重庆市自然科学基金重点研发项目(cstc2020jcyj-zdxmX0005);重庆市市级财政资金项目(01010106)。

摘　　要：长链非编码RNA(long noncoding RNA,LncRNA)是一类长度大于200个核苷酸且不能编码蛋白质的RNA,可通过多种调控途径作用于皮肤恶性黑色素瘤的增殖、侵袭和转移等。为探究酉州乌羊(Capra hircus)LncRNA TCONS_00153149基因在黑色素沉积过程的调控作用及分子机制,本研究以酉州乌羊和渝东白山羊为对象,验证LncRNA TCONS_00153149在不同肤色山羊组织的表达特性,同时分析其在小鼠(Mus musculus)B16-F10黑色素细胞分化阶段的表达特征,通过构建LncRNA TCONS_00153149的慢病毒表达载体,转染B16-F10黑色素细胞,探究LncRNA TCONS_00153149对黑色素沉积的调控作用。结果显示LncRNA TCONS_00153149在酉州乌羊和渝东白山羊不同组织中均有不同程度的表达,且酉州乌羊皮肤组织中的相对表达量极显著高于渝东白山羊(P<0.01);LncRNA TCONS_00153149的表达在黑色素细胞分化前期呈上升趋势,分化第3天相对表达量达到最高,其后呈递减趋势,在分化第7天达到最低,其在第3天的相对表达量极显著高于分化第0、1和7天(P<0.01);转染B16-F10细胞发现,与转空载体组(negative control group,NC)相比,转染LncRNA TCONS_00153149基因的黑色素细胞中,LncRNA TCONS_00153149基因相对表达量极显著提高(P<0.01),黑色素含量显著升高,这些结果验证了LncRNA TCONS_00153149基因对黑色素沉积的显著促进作用。同时,黑色素沉积相关基因酪氨酸酶相关蛋白1(TYRosinase related protein 1,TYRP1)、小眼畸形转录因子(microphthalmia-associated transcription factor,MITF)、酪氨酸酶相关蛋白2基因(TYRosinase-related protein 2,TYRP2)相对表达量显著升高(P<0.05),Agouti信号蛋白(agouti signaling protein,ASIP)相对表达量极显著降低(P<0.01)。结合Inta RNA2.0.0软件分析LncRNA TCONS_00153149与色素关键基因的互作结果,本研究推测LncRNA TCONS_00153149基因可能是通过靶向增强黑色素候选基因(TYRP1,TYRP2,MITF,ASIP基因等)的功能,正向调控黑�LncRNA was widely transcribed in the mammalian genome and promote the proliferation,invasion and metastasis of skin malignant melanoma through a variety of regulatory pathways.To further explore the effect of LncRNA TCONS_00153149 gene on the process of melanin deposition in Youzhou Black goat(Capra hircus),the tissue expression profile of LncRNA TCONS_00153149 were detected using qPCR method in different tissues of Youzhou dark goat and Yudong white goat,as well as the LncRNA TCONS_00153149 expression characteristics were detected in B16-F10 cells at differentiation stage stages,the LncRNA TCONS_00153149 lentiviral vector was constructed and transfected into B16-F10 melanocytes to explore the effect on the process of melanin deposition.The results demonstrated that LncRNA TCONS_00153149 was widely expressed in different tissues of the goat,especially with significantly higher expression in the skin in Youzhou black goat than that of Yudong white goat(P<0.01).Comparison analysis illustrated that the expression pattern of the LncRNA TCONS_00153149 was increasing in the early stage of melanocyte differentiation,and reached the highest relative expression on the 3rd day,and then decreasing after that,the expression was the lowest on the 7th day.The expression on the 3rd day was significantly higher than that of the 0,1,and 7 days(P<0.01),showed that LncRNA TCONS_00153149 have a positive regulatory relationship to melanin deposition.The transfection experiment found that the relative expression of LncRNA TCONS_00153149 gene was extremely significantly increased(P<0.01),and the melanin content was significantly increased compared with the negative control group(NC),these results verified the significant promotion effect of LncRNA TCONS_00153149 gene on melanin deposition.Besides,The relative expression of TYRosinase related protein 1(TYRP1),microphthalmia-associated transcription factor(MITF),TYRosinase-related protein 2(TYRP2)was significantly increased(P<0.05),and the relative expression of agouti signaling protein(

关 键 词：长链非编码RNA(LncRNA) 黑色素沉积 乌质性状 酉州乌羊 

分 类 号：S821.2[农业科学—畜牧学]