Cloning and Tissue Expression Profile of KLF9 Gene in Goat

doi:10.7668/hbnxb.2017.06.016

Abstract

Abstract: The aim of this study was to clone the sequence of Kruppel-like factor 9 (KLF9) gene and elucidate its expression in different tissues and cells in goats,which may lay a foundation for further study of the KLF9 gene in regulating goat intramuscular fat deposition.Two 7-day-old goats were used for intramuscular preadipocytes culture by collagenase digestion.Various tissues,including heart,liver,spleen,kidney,longissimus dorsi muscle,biceps femoris muscle,triceps brachii muscle and subcutaneous fat,were collected from 4 adult Jianzhou Big-eared goats and Tibetan goats respectively.After total RNA extraction,liver,spleen and subcutaneous fat tissues were used for KLF9 gene cloning by RT-PCR.Real-time fluorescence quantitative PCR (qPCR) method was then used to detect the expression level of KLF9 gene in different tissues and intramuscular pre-adipocytes of different differentiation stages(0,2,4,6 d).The results showed that the length of goat KLF9 gene was 891 bp,including 735 bp CDS,86 bp 5'UTR sequence and 70 bp 3'UTR sequence,encoding 244 amino acids.The KLF9 amino acid sequences of Jianzhou Big-eared goats shared 99.73% similarity with Tibetan goats, following by cattle,pig,human and rat with 99.18%-97.54%,respectively.The expression level of KLF9 gene in liver and subcutaneous fat of Jianzhou Big-eared goats was significantly higher than that in other tissues (P <0.01),while the expression level of KLF9 gene in lungs of Tibetan goats was significantly higher than that in any other tissues(P <0.01).Comparing with the non-differentiation pre-adipocytes,the expression of KLF9 gene was significantly increased in 2 d(P <0.01).The expression pattern of the gene was specific in varieties on the basis of obtaining the KLF9 gene sequence of goat,and it might play an important role in the early stage of differentiation.

Key words: Goat, KLF9, Cloning, Tissue expression, Intramuscular preadipocytes

CLC Number:

Q78
S826

ZHAO Yue, LIN Yaqiu, ZHU Jiangjiang, LIN Sen, CHI Yongdong, LIU Lushu, WANG Yong. Cloning and Tissue Expression Profile of KLF9 Gene in Goat[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2017, 32(6): 106-112. doi: 10.7668/hbnxb.2017.06.016.

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References

[1] 胡雅淇,林海,李军.中国城乡居民羊肉消费差异分析[J].农业展望,2017,13(1):77-81.
[2] 杜琛.绒山羊肌内脂肪沉积特征及脂肪细胞分化机理的研究[D].呼和浩特:内蒙古农业大学,2014.
[3] Huang Z G,Xiong L,Liu Z S,et al.The developmental changes and effect on IMF content of H-FABP and PPARgamma mRNA expression in sheep muscle[J].Acta Genetica Sinica,2006,33(6):507-514.
[4] 栾兆进,曲绪仙,贺建宁,等.羊肌内脂肪调控研究进展[J].家畜生态学报,2014(9):8-13.
[5] Gao Y,Cao Q,Lu L,et al.Kruppel-like factor family genes are expressed during Xenopus embryogenesis and involved in germ layer formation and body axis patterning[J].Developmental Dynamics:an Official Publication of the American Association of Anatomists,2015,244(10):1328-1346.
[6] Pei J,Grishin N V.A new family of predicted Krüppel-like factor genes and pseudogenes in placental mammals[J].PLoS One,2013,8(11):e81109.
[7] Bieker J J.Kruppel-like factors:three fingers in many pies[J].Journal of Biological Chemistry,2001,276(37):34355-34358.
[8] Swamynathan S K.Krüppel-like factors:three fingers in control[J].Human Genomics,2010,4(4):1-8.
[9] Wu Z,Wang S.Role of kruppel-like transcription factors in adipogenesis[J].Developmental Biology,2013,373(2):235-243.
[10] Birsoy K,Chen Z,Friedman J.Transcriptional regulation of adipogenesis by KLF4[J].Cell Metabolism,2008,7(4):339-347.
[11] Li D,Yea S,Li S,et al.Krüppel-like factor-6 promotes preadipocyte differentiation through histone deacetylase 3-dependent repression of DLK1[J].The Journal of Biological Chemistry,2005,280(29):26941-26952.
[12] Lee H,Kim H J,Lee Y J,et al.Krüppel-like factor KLF8 plays a critical role in adipocyte differentiation[J].PLoS One,2012,7(12):e52474.
[13] Pei H,Yao Y,Yang Y,et al.Krüppel-like factor KLF9 regulates PPARγ transactivation at the middle stage of adipogenesis[J].Cell Death and Differentiation,2011,18(2):315-327.
[14] Banerjee S S,Feinberg M W,Watanabe M,et al.The krüppel-like factor KLF2 inhibits peroxisome proliferator-activated receptor-gamma expression and adipogenesis[J].The Journal of Biological Chemistry,2003,278(4):2581-2584.
[15] Sue N,Jack B H,Eaton S A,et al.Targeted disruption of the basic Krüppel-like factor gene (Klf3) reveals a role in adipogenesis[J].Molecular and Cellular Biology,2008,28(12):3967-3978.
[16] Jiang S,Wei H,Song T,et al.KLF13 promotes porcine adipocyte differentiation through PPARγ activation[J].Cell & Bioscience,2015,5(1):28.
[17] Imataka H,Sogawa K,Yasumoto K,et al.Two regulatory proteins that bind to the basic transcription element (BTE),a GC box sequence in the promoter region of the rat P-4501A1 gene[J].The EMBO Journal,1992,11(10):3663-3671.
[18] Shen P,Sun J,Xu G,et al.KLF9,a transcription factor induced in flutamide-caused cell apoptosis,inhibits AKT activation and suppresses tumor growth of prostate cancer cells[J].The Prostate,2014,74(9):946-958.
[19] Simmen F A,Su Y,Xiao R,et al.The krüppel-like factor 9(KLF9) network in HEC-1-A endometrial carcinoma cells suggests the carcinogenic potential of dys-regulated KLF9 expression[J].Reproductive Biology and Endocrinology,2008,6(1):41.
[20] Li J Z,Li J,Wang H Q,et al.MiR-141-3p promotes prostate cancer cell proliferation through inhibiting kruppel-like factor-9 expression[J].Biochemical and Biophysical Research Communications,2017,482(4):1381-1386.
[21] Qiao F,Yao F,Chen L,et al.Krüppel-like factor 9 was down-regulated in esophageal squamous cell carcinoma and negatively regulated beta-catenin/TCF signaling[J].Molecular Carcinogenesis,2016,55(3):280-291.
[22] Ying M,Tilghman J,Wei Y,et al.Kruppel-like factor-9(KLF9) inhibits glioblastoma stemness through global transcription repression and integrin α6 inhibition[J].The Journal of Biological Chemistry,2014,289(47):32742-32756.
[23] Fu D Z,Cheng Y,He H,et al.The fate of Krüppel-like factor 9-positive hepatic carcinoma cells may be determined by the programmed cell death protein 5[J].International Journal of Oncology,2014,44(1):153-160.
[24] Pabona J M,Simmen F A,Nikiforov M A,et al.Krüppel-like factor 9 and progesterone receptor coregulation of decidualizing endometrial stromal cells:implications for the pathogenesis of endometriosis[J].The Journal of Clinical Endocrinology and Metabolism,2012,97(3):376-392.
[25] Pabona J P,Velarde M C,Zeng Z,et al.Nuclear receptor co-regulator Krüppel-like factor 9 and prohibitin 2 expression in oestrogen-induced epithelial cell proliferation in the mouse uterus[J].Journal of Endocrinology,2009,200(1):63-73.
[26] Sunadome K,Yamamoto T,Ebisuya M,et al.ERK5 regulates muscle cell fusion through Klf transcription factors[J].Developmental Cell,2011,20(2):192-205.
[27] Kimura H,Fujimori K.Activation of early phase of adipogenesis through Krüppel-like factor KLF9-mediated,enhanced expression of CCAAT/enhancer-binding protein β in 3T3-L1 cells[J].Gene,2014,534(2):169-176.
[28] Jelinic P,Schlappe B A,Conlon N,et al.Concomitant loss of SMARCA2 and SMARCA4 expression in small cell carcinoma of the ovary,hypercalcemic type[J].Modern Pathology:an Official Journal of the United States and Canadian Academy of Pathology,Inc,2016,29(1):60-66.
[29] Vangamudi B,Paul T A,Shah P K,et al.The SMARCA2/4 ATPase domain surpasses the bromodomain as a drug target in SWI/SNF-Mutant cancers:insights from cDNA rescue and PFI-3 inhibitor studies[J].Cancer Research,2015,75(18):3865-3878.
[30] Tan C,Xuan L,Cao S,et al.Decreased histone deacetylase 2(HDAC2) in peripheral blood monocytes (PBMCs) of COPD patients[J].PLoS One,2016,11(1):e0147380.
[31] Fei H J,Zu L D,Wu J,et al.PCAF acts as a gastric cancer suppressor through a novel PCAF-p16-CDK4 axis[J].American Journal of Cancer Research,2016,6(12):2772-2786.
[32] 焦涛.KLF9通过诱导PGC-1α调节小鼠原代肝脏细胞抗ROS基因的表达[D].北京:北京协和医学院,2014.
[33] 王翠喆,哈晓丹,谢建新,等.KLF9和KLF15在维吾尔族网膜脂肪中的表达及其与炎症相关性分析[J].中国免疫学杂志,2016,32(11):1653-1656,1660.
[34] Antin P B,Pier M,Sesepasara T,et al.Embryonic expression of the chicken Krüppel-like (KLF) transcription factor gene family[J].Developmental Dynamics:an Official Publication of the American Association of Anatomists,2010,239(6):1879-1887.
[35] Martin K M,Metcalfe J C,Kemp P R.Expression of Klf9 and Klf13 in mouse development[J].Mechanisms of Development,2001,103(1/2):149-151.
[36] Mori T,Sakaue H,Iguchi H,et al.Role of krüppel-like factor 15(KLF15) in transcriptional regulation of adipogenesis[J].The Journal of Biological Chemistry,2005,280(13):12867-12875.

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