STAT3基因对广西黄牛肌肉干细胞成肌诱导分化的调控研究  

Study on Regulation of STAT 3 Gene on Myogenic Differentiation of Muscle Stem Cells in Guangxi Cattle

作　　者：吕巧 邹超霞 吴永梅 张瑞门 郑自华[2] 石德顺[1] 韦英明[2] 邓彦飞[1] LYU Qiao;ZOU Chaoxia;WU Yongmei;ZHANG Ruimen;ZHENG Zihua;SHI Deshun;WEI Yingming;DENG Yanfei(State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources,College of Animal Science and Technology,Guangxi University,Nanning 530004,China;Research Institute of Agriculture and Animal Husbandry Industry Development,Guangxi University,Nanning 530004,China)

机构地区：[1]广西大学动物科学技术学院,亚热带生物资源保护利用国家重点实验室,南宁530004 [2]广西大学农牧产业发展研究院,南宁530004

出　　处：《中国畜牧兽医》2021年第8期2771-2777,共7页China Animal Husbandry & Veterinary Medicine

基　　金：国家现代农业产业技术体系广西创新团队建设项目(nycytxgxcxtd-09-01);广西自然科学基金(桂科AA17204051、2018GXNSFAA281007);南宁市科学技术局项目(20194147、20192087);畜禽生态养殖模式及关键技术创新示范(桂科AA17204052);大石山区山羊主要疫病综合防控关键共性技术研发与应用示范(桂科AB19245010)。

摘　　要：研究旨在检测信号转导与转录激活因子3(STAT3)的表达规律,克隆STAT 3基因,构建真核表达载体,并探索STAT 3基因对广西黄牛肌肉干细胞(MuSCs)分化的调控作用。采集6、12、18月龄广西黄牛肌肉组织以及生长期(GM)和分化期(DM)肌肉干细胞,分别提取RNA并反转录为cDNA,通过实时荧光定量PCR检测STAT 3基因和成肌相关基因的表达规律;克隆黄牛STAT 3基因完整编码区序列,构建过表达载体pCD-STAT3并检测其对黄牛肌肉干细胞分化的影响。实时荧光定量PCR结果表明,STAT 3基因在6、12、18月龄黄牛肌肉组织中均有表达,且在18月龄中表达量最高,12月龄中表达量最低;STAT 3和成肌调节因子6(MYF 6)基因在分化期肌肉干细胞中表达量均极显著高于生长期(P<0.01)。广西黄牛STAT 3基因编码区全长2313 bp,与空载体pCDNA3.1相连成功构建过表达载体pCD-STAT3,将pCD-STAT3转入体外培养广西黄牛肌肉干细胞,肌肉干细胞中STAT 3基因及成肌决定蛋白(MyoD1)、骨骼肌肌球蛋白重链(MyHC)基因表达量极显著升高(P<0.01),肌细胞生成素(MyoG)基因表达量显著升高(P<0.05),且肌管数量、大小均显著高于pCDNA3.1组(P<0.05)。本研究检测了转录因子STAT3在广西黄牛背最长肌中的表达规律;且过表达STAT 3基因显著促进了广西黄牛肌肉干细胞的成肌分化,为深入研究STAT 3基因在肌肉生长发育中的作用及其分子调控机制奠定基础。This study was aimed to detect the expression pattern of signal transduction and activator of transcription 3(STAT3),clone STAT 3 gene,construct eukaryotic expression vector,and explore the regulatory effect of STAT 3 gene on the differentiation of muscle stem cells in Guangxi cattle.Muscle tissues of Guangxi cattle aged 6,12 and 18 months,GM and DM stages muscle stem cells were collected,and RNA was extracted and reverse transcribed into cDNA.The expression pattern of STAT 3 gene was detected by Real-time quantitative PCR,and the complete coding region sequence of STAT 3 gene was amplified to construct over-expression vector pCD-STAT3,so as to test its effects on the differentiation of cattle muscle stem cells.The results of Real-time quantitative PCR showed that STAT 3 gene expressed in the muscle tissues of cattle aged at 6,12 and 18 months,and the expression level was the highest at the age of 18 months and the lowest at 12 months.The expression levels of STAT 3 and myogenic regulatory factor 6(MYF6)genes in differentiated MSCs were extremely significantly higher than those in proliferative MSCs(P<0.01).The coding region of STAT 3 gene in Guangxi cattle was 2313 bp,which was successfully linked with empty vector pcDNA3.1 to construct the overexpression vector pCD-STAT3.The pCD-STAT3 was transferred into muscle stem cells in Guangxi cattle in vitro,the expression of STAT 3 gene and myoblast differentiation marker genes myogenic differentiation 1(MyoD1)and muscle myosin heavy chain(MyHC)in muscle stem cells were extremely significantly increased(P<0.01),and the expression of myopoietin(MyoG)gene was significantly increased(P<0.05).Moreover,the number and size of myotubes in pCD-STAT3 group were significantly higher than those in pcDNA3.1 group(P<0.05).In this study,the expression pattern of STAT3 in longissimus dorsi muscle of Guangxi cattle was investigated,and overexpression of STAT 3 gene could significantly promote the differentiation of bovine muscle stem cells,which laid a foundation for further study on

关 键 词：广西黄牛 转录因子 STAT3基因 肌肉干细胞 

分 类 号：Q254[生物学—细胞生物学]