STAT3基因对广西黄牛肌肉干细胞成肌诱导分化的调控研究

doi:10.16431/j.cnki.1671-7236.2021.08.010

Abstract

Abstract: This study was aimed to detect the expression pattern of signal transduction and activator of transcription 3 (STAT3), clone STAT3 gene, construct eukaryotic expression vector, and explore the regulatory effect of STAT3 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 STAT3 gene was detected by Real-time quantitative PCR, and the complete coding region sequence of STAT3 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 STAT3 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 STAT3 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 STAT3 gene in Guangxi cattle was 2 313 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 STAT3 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 STAT3 gene could significantly promote the differentiation of bovine muscle stem cells, which laid a foundation for further study on the role of STAT3 gene in muscle growth and development and its molecular regulatory mechanism.

Key words: Guangxi cattle; transcription factor; STAT3 gene; muscle stem cell

CLC Number:

Q254

LYU Qiao, ZOU Chaoxia, WU Yongmei, ZHANG Ruimen, ZHENG Zihua, SHI Deshun, WEI Yingming, DENG Yanfei. Study on Regulation of STAT3 Gene on Myogenic Differentiation of Muscle Stem Cells in Guangxi Cattle[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(8): 2771-2777.

References

[1] HOU J S U,HENZEL W J,HO T C,et al.An interleukin-4-induced transcription factor:IL-4 Stat[J].Science,1994,265(5179):1701-1706.
[2] ZHONG Z W Z,DARNELL J E J R.Stat3:A STAT family member activated by tyrosine phosphorylation in response to epidermal growth factor and interleukin-6[J].Science,1994,264(5155):95.
[3] JIANG L,ZHANG J,WANG J J,et al.Sperm,but not oocyte,DNA methylome is inherited by zebrafish early embryos[J].Cell,2013,153(4):773-784.
[4] 钟忆.STAT3乙酰化在白介素6调控谷氨酸诱导神经细胞凋亡中的作用[D].遵义:遵义医科大学,2020. ZHONG Y.The role of STAT3 acetylation in the regulation of glutamate-induced neuronal cells apoptosis by interleukin-6[D].Zunyi:Zunyi Medical University,2020.(in Chinese)
[5] TAKEDA K N K,SHI W,ET A L.Targeted disruption of the mouse STAT3 gene leads to early embryonic lethality[J].Proceedings National Academy of Sciences of the United States of America,1997,94(8):3801-3804.
[6] TENG Y R J L,COWELL J K.The involvement of JAK-STAT3 in cell motility,invasion,and metastasis[J].JAKSTAT,2014,3(1):e28086.
[7] HUGHES K,WATSON C.The multifaceted role of STAT3 in mammary gland involution and breast cancer[J].International Journal of Molecular Sciences,2018,19(6):1695.
[8] KASEMBELI M,BHARADWAJ U,ROBINSON P,et al.Contribution of STAT3 to inflammatory and fibrotic diseases and prospects for its targeting for treatment[J].International Journal of Molecular Sciences,2018,19(8):2299.
[9] AVALLE L,CAMPOREALE A,CAMPERI A,et al.STAT3 in cancer:A double edged sword[J].Cytokine,2017,98:42-50.
[10] NUNES A M,WUEBBLES R D,SARATHY A,et al.Impaired fetal muscle development and JAK-STAT activation mark disease onset and progression in a mouse model for merosin-deficient congenital muscular dystrophy[J].Human Molecular Genetics,2017,26(11):2018-2033.
[11] GUADAGNIN E,NAROLA J,BÖNNEMANN C G,et al.Tyrosine 705 phosphorylation of STAT3 is associated with phenotype severity in TGF-β1 transgenic mice[J].BioMed Research International,2015,2015:843743.
[12] WADA E,TANIHATA J,IWAMURA A,et al.Treatment with the anti-IL-6 receptor antibody attenuates muscular dystrophy via promoting skeletal muscle regeneration in dystrophin-/utrophin-deficient mice[J].Skeletal Muscle,2017,7(1):23.
[13] BENITO C,DAVIS C M,GOMEZ-SANCHEZ J A,et al.STAT3 controls the long-term survival and phenotype of repair Schwann cells during nerve regeneration[J].The Journal of Neuroscience,2017,37(16):4255-4269.
[14] LIAO X H,XIANG Y,LI H,et al.VEGF-A stimulates STAT3 activity via nitrosylation of myocardin to regulate the expression of vascular smooth muscle cell differentiation markers[J].Scientific Reports,2017,7(1):2660.
[15] YANG X,LIN A,JIANG N,et al.Interleukin-6 trans-signalling induces vascular endothelial growth factor synthesis partly via Janus kinases-STAT3 pathway in human mesothelial cells[J].Nephrology,2017,22(2):150-158.
[16] SU Y L,BANERJEE S,WHITE S,et al.STAT3 in tumor-associated myeloid cells:Multitasking to disrupt immunity[J].International Journal of Molecular Sciences,2018,19(6):1803.
[17] YANG X,WANG Y,SUN X,et al.STAT3 activation is associated with interleukin-10 expression and survival in primary central nervous system lymphoma[J].World Neurosurge,2020,134:e1077-e1084.
[18] QIN J J,YAN L,ZHANG J,et al.STAT3 as a potential therapeutic target in triple negative breast cancer:A systematic review[J].Journal of Experimental & Clinical Cancer Rearch,2019,38(1):195.
[19] XU S,NI H,CHEN H,et al.The interaction between STAT3 and nAChRα1 interferes with nicotine-induced atherosclerosis via Akt/mTOR signaling cascade[J].Aging,2019,11(19):8120-8138.
[20] 吴贤锋.山羊STAT3和STAT5A基因SNPs、mRNA表达及甲基化对生产性能的影响[D].杨凌:西北农林科技大学,2015. WU X F.SNPs,mRNA expression and DNA methylation of STAT3 and STAT5A genes and their influences on production traits in goats[D].Yangling:Northwest A&F University,2015.(in Chinese)
[21] WANG J,YANG L Z,ZHANG J S,et al.Effects of microRNAs on skeletal muscle development[J].Gene,2018,668:107-113.
[22] TIERNEY M T,AYDOGDU T,SALA D,et al.STAT3 signaling controls satellite cell expansion and skeletal muscle repair[J].Nature Medicine,2014,20(10):1182-1186.

Study on Regulation of STAT3 Gene on Myogenic Differentiation of Muscle Stem Cells in Guangxi Cattle

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