miR-10b对延边黄牛垂体细胞中GH分泌的影响  

Effect of miR-10b on GH secretion in pituitary cells of Yanbian yellow cattle

作　　者：娄安钢[1] 季久秀 相思宇 金太花 陈莹莹[1] 张睿[1] 崔长艳 于龙政[1] 关立增[1,2] LOU An-gang;JI Jiu-xiu;XIANG Si-yu;JIN Tai-hua;CHEN Ying-ying;ZHANG Rui;CUI Chang-yan;YU Long-zheng;GUAN Li-zeng(College of Agriculture,Yanbian University,Yanji,Jilin 133002,China;College of Agriculture and Forestry Sciences,Linyi University,Li-nyi,Shandong 276005,China)

机构地区：[1]延边大学农学院,吉林延吉133002 [2]临沂大学农林科学学院,山东临沂276005

出　　处：《中国兽医学报》2020年第8期1641-1646,共6页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31660614)。

摘　　要：选择在延边黄牛和韩延牛中显著差异表达的miR-10b,利用实时定量PCR(qPCR)和Western blot技术,研究miR-10b对延边黄牛垂体细胞中GH分泌水平的影响及miR-10b与靶基因之间的调控机制。qPCR和Western blot结果显示,miR-10b水平的增加能显著提高延边黄牛垂体细胞中GH mRNA和蛋白的表达(P<0.01),降低miR-10b水平可抑制GH mRNA和蛋白的表达。生物信息学分析和双荧光素酶报告基因系统结果显示,miR-10b靶向了SSTR2的3′UTR;miR-10b水平的增加能够显著抑制SSTR2 mRNA和蛋白的表达(P<0.01),反之降低miR-10b水平能显著提高SSTR2 mRNA和蛋白的表达(P<0.01)。结果表明,miR-10b可通过调节SSTR2基因的表达而调控垂体细胞中GH的分泌,这一结果为研究miRNA调控动物生长发育机制提供了理论依据。In order to study the effect of miRNAs on GH secretion in pituitary cells of Yanbian yellow cattle,Yanbian yellow cattle and Han Yan cattle was selected to study the effect of the differently expressed miR-10 b on GH secretion in pituitary cells of Yanbian yellow cattle and the regulation mechanism between miR-10 b and target genes using real-time quantitative PCR(qPCR)and Western blot in this study.The results qPCR and Western blot showed that the expression of GH mRNA and protein in pituitary cells of Yanbian yellow cattle could be significantly increased by increasing the level of miR-10 b(P<0.01),whereas the decrease of the level of miR-10 b could inhibit the expression of GH mRNA and protein.The results of bioinformatics analysis and double luciferase reporter gene system validation showed that miR-10 b targeted 3′UTR of SSTR2.The increase of the level of miR-10 b could significantly inhibit the expression of SSTR2 mRNA and protein(P<0.01),whereas the decrease of the level of miR-10 b could significantly increase the expression of SSTR2 mRNA and protein(P<0.01).This study proved that miR-10 b could regulate GH secretion in pituitary cells by regulating the expression of SSTR2 gene.The results of this study will provide a theoretical basis for studying the mechanism of growth and development of animals regulated by miRNAs.

关 键 词：MIR-10B 延边黄牛 垂体细胞 GH SSTR2 

分 类 号：S813[农业科学—畜牧学] S852.2[农业科学—畜牧兽医]