过表达circNMT1促进水牛脂肪细胞的成脂分化

doi:10.11843/j.issn.0366-6964.2022.04.008

Abstract

Abstract: The aim of this research was to identify non-coding RNA, circNMT1, clarify its tissue and cells expression patterns, and explore the effect of circNMT1 overexpression on adipocyte differentiation. In the study, the heart, liver, spleen, lung, kidney, longissimus dorsi, dorsal subcutaneous adipose tissue and preadipocytes of 30-month-old Chinese swamp buffalo (Xinyang buffalo, n=3) and 3T3-L1 cells were used as experimental materials. Semiquantitative PCR and real-time quantitative PCR (qRT-PCR) were used to identify circNMT1, explore its location in the nucleus or cytoplasm of buffalo adipocytes. Relative expression level of circNMT1 in different tissues and preadipocytes of buffalo at different differentiation stages were detected by qRT-PCR. circNMT1 was overexpressed in 3T3-L1 cells and buffalo preadipocytes respectively, and the accumulation of lipid droplet was detected by morphology and quantitative method. Meanwhile, the relative expression level of adipogenic differentiation marker genes was detected by qRT-PCR. The results showed that circNMT1 was stably expressed circRNA, which was expressed in both nucleus and cytoplasm of buffalo preadipocytes. And circNMT1 was highly expressed in adipose tissue and maturation adipocytes (P<0.001). Gain-of-function experiments demonstrated that the overexpression of circNMT1 significantly promoted the accumulation of lipid droplets in 3T3-L1 cells and buffalo adipocytes, and significantly upregulated the relative expression level of adipogenic marker genes PPARG, C/EBPα and FABP4 (P<0.01). These results indicated that circNMT1 might be a positive regulator for buffalo adipocytes differentiation, which would provide new insights into the regulatory roles of circNMT1 in buffalo adipocytes.

Key words: buffalo, IMF, adipocytes, circRNA, overexpression

CLC Number:

S823.83

FENG Xue, ZHAO Jinhui, WANG Shuzhe, HUANG Jieping, WEI Xuefeng, SHI Yuangang, MA Yun. Overexpression of circNMT1 Promotes Adipogenic Differentiation of Buffalo Adipocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(4): 1077-1088.

References 46

[1]	PINEDA P S,FLORES E B,HERRERA J R V,et al.Opportunities and challenges for improving the productivity of swamp buffaloes in southeastern Asia[J].Front Genet, 2021,12:629861.
[2]	LOWE C E,O’RAHILLY S,ROCHFORD J J.Adipogenesis at a glance[J].J Cell Sci, 2011,124(16):2681-2686.
[3]	岳永起,华永琳,熊燕,等.microRNA调控动物皮下脂肪组织和肌内脂肪沉积的研究进展[J].畜牧兽医学报,2021,52(10):2698-2709.YUE Y Q,HUA Y L,XIONG Y,et al.Research progress of microRNA regulation on the fat deposition of subcutaneous adipose tissue and intramuscular fat in animals[J].Acta Veterinaria et Zootechnica Sinica, 2021,52(10):2698-2709.(in Chinese)
[4]	LEI Z X,WU H G,XIONG Y,et al.ncRNAs regulate bovine adipose tissue deposition[J].Mol Cell Biochem, 2021,476(7): 2837-2845.
[5]	HUANG J P,ZHENG Q Z,WANG S Z,et al.High-throughput RNA sequencing reveals NDUFC 2-AS lncRNA Promotes adipogenic differentiation in Chinese buffalo (Bubalus bubalis L)[J].Genes, 2019,10(9):689.
[6]	ZHANG Y F,GUO X,PEI J,et al.CircRNA expression profile during yak adipocyte differentiation and screen potential circRNAs for adipocyte differentiation[J].Genes, 2020,11(4):414.
[7]	ZHANG M,HAN Y,ZHAI Y H,et al.Integrative analysis of circRNAs,miRNAs,and mRNAs profiles to reveal ceRNAs networks in chicken intramuscular and abdominal adipogenesis[J].BMC Genomics, 2020,21(1):594.
[8]	WANG L D,LIANG W S,WANG S S,et al.Circular RNA expression profiling reveals that circ-PLXNA1 functions in duck adipocyte differentiation[J].PLoS One, 2020,15(7):e0236069.
[9]	LI A,HUANG W L,ZHANG X X,et al.Identification and characterization of circRNAs of two pig breeds as a new biomarker in metabolism-related diseases[J].Cell Physiol Biochem, 2018,47(6):2458-2470.
[10]	LIU X,LIU K Q,SHAN B S,et al.A genome-wide landscape of mRNAs,lncRNAs,and circRNAs during subcutaneous adipogenesis in pigs[J].J Anim Sci Biotechnol, 2018,9:76.
[11]	王卫振,邓占钊,辛国省,等.环状RNA的生物学功能及其在家禽中的研究进展[J].畜牧兽医学报,2021,52(7):1778-1788.WANG W Z,DENG Z Z,XIN G S,et al.The biological function of circular RNA and its research progress in poultry[J].Acta Veterinaria et Zootechnica Sinica, 2021,52(7):1778-1788.(in Chinese)
[12]	LIANG W C,WONG C W,LIANG P P,et al.Translation of the circular RNA circβ-catenin promotes liver cancer cell growth through activation of the Wnt pathway[J].Genome Biol, 2019,20(1):84.
[13]	WEI X F,LI H,YANG J M,et al.Circular RNA profiling reveals an abundant circLMO7 that regulates myoblasts differentiation and survival by sponging miR-378a-3p[J].Cell Death Dis, 2017,8(10):e3153.
[14]	MEMCZAK S,JENS M,ELEFSINIOTI A,et al.Circular RNAs are a large class of animal RNAs with regulatory potency[J].Nature, 2013,495(7441):333-338.
[15]	LIU J Y,ZHANG C H,ZHANG B Y,et al.Comprehensive analysis of the characteristics and differences in adult and newborn brown adipose tissue (BAT):newborn BAT is a more active/dynamic BAT[J].Cells, 2020,9(1):201.
[16]	LONG T,GUO Z Y,HAN L,et al.Differential expression profiles of circular RNAs during osteogenic differentiation of mouse adipose-derived stromal cells[J].Calcif Tissue Int, 2018,103(3):338-352.
[17]	HUANG J P,ZHAO J H,ZHENG Q Z,et al.Characterization of circular RNAs in Chinese buffalo (Bubalus bubalis ) adipose tissue:a focus on circular RNAs involved in fat deposition[J].Animals, 2019,9(7):403.
[18]	冯雪.水牛肌内脂肪沉积关键基因的筛选及相关功能研究[D].信阳:信阳师范学院,2021.FENG X.Screening and functional analysis of important genes for intramuscular fat deposition in buffaloes[D].Xinyang: Xinyang Normal University,2021.(in Chinese)
[19]	ZHU R R,FENG X,WEI Y T,et al.lncSAMM50 enhances adipogenic differentiation of buffalo adipocytes with no effect on its host gene[J].Front Genet, 2021,12:626158.
[20]	冯雪,洪慧颖,孙晓菲,等.水牛PPARG来源环状RNA的鉴定及表达谱分析[J].中国畜牧兽医,2019,46(12):3457-3465.FENG X,HONG H Y,SUN X F,et al.Identification and expression profile analysis of PPARG-derived circular RNA in buffalo[J].China Animal Husbandry & Veterinary Medicine, 2019,46(12):3457-3465.(in Chinese)
[21]	WRIGHT M H,HEAL W P,MANN D J,et al.Protein myristoylation in health and disease[J].J Chem Biol, 2010,3(1):19-35.
[22]	XU X L,ZHANG J W,TIAN Y H,et al.CircRNA inhibits DNA damage repair by interacting with host gene[J].Mol Cancer, 2020,19(1):128.
[23]	DING Z Y,SUN D Y,HAN J,et al.Novel noncoding RNA CircPTK2 regulates lipolysis and adipogenesis in cachexia[J].Mol Metab, 2021,53:101310.
[24]	WANG L,ZHOU J H,ZHANG C,et al.A novel tumour suppressor protein encoded by circMAPK14 inhibits progression and metastasis of colorectal cancer by competitively binding to MKK6[J].Clin Transl Med, 2021,11(10):e613.
[25]	CHEN W B,CEN S R,ZHOU X M,et al.Circular RNA CircNOLC1,upregulated by NF-KappaB,promotes the progression of prostate cancer via miR-647/PAQR4 axis[J].Front Cell Dev Biol, 2021,8:624764.
[26]	HANSEN T B,JENSEN T I,CLAUSEN B H,et al.Natural RNA circles function as efficient microRNA sponges[J]. Nature, 2013,495(7441):384-388.
[27]	LI Z Y,HUANG C,BAO C,et al.Exon-intron circular RNAs regulate transcription in the nucleus[J].Nat Struct Mol Biol, 2015, 22(3):256-264.
[28]	JECK W R,SORRENTINO J A,WANG K,et al.Circular RNAs are abundant,conserved,and associated with ALU repeats[J]. RNA, 2013,19(2):141-157.
[29]	WANG P L,BAO Y,YEE M C,et al.Circular RNA is expressed across the eukaryotic tree of life[J].PLoS One, 2014,9(6): e90859.
[30]	XIAO W,LI J,HU J,et al.Circular RNAs in cell cycle regulation:mechanisms to clinical significance[J].Cell Prolif, 2021: e13143,doi:10.1111/cpr.13143.
[31]	WEI X F,HAN S,WANG S Z,et al.ANGPTL8 regulates adipocytes differentiation and adipogenesis in bovine[J].Gene, 2019,707:93-99.
[32]	BOSCH L,TOR M,REIXACH J,et al.Age-related changes in intramuscular and subcutaneous fat content and fatty acid composition in growing pigs using longitudinal data[J].Meat Sci, 2012,91(3):358-363.
[33]	TYRA M,ROPKA-MOLIK K,TERMAN A,et al.Association between subcutaneous and intramuscular fat content in porcine ham and loin depending on age,breed and FABP 3 and LEPR genes transcript abundance[J].Mol Biol Rep, 2013,40(3):2301-2308.
[34]	HUDSON N J,REVERTER A,GRIFFITHS W J,et al.Gene expression identifies metabolic and functional differences between intramuscular and subcutaneous adipocytes in cattle[J].BMC Genomics, 2020,21(1):77.
[35]	YAMADA T,KAMIYA M,HIGUCHI M.Fat depot-specific effects of body fat distribution and adipocyte size on intramuscular fat accumulation in Wagyu cattle[J].Anim Sci J, 2020,91(1):e13449.
[36]	CAMPOS C F,DUARTE M S,GUIMARAES S E F,et al.Review:animal model and the current understanding of molecule dynamics of adipogenesis[J].Animal, 2016,10(6):927-932.
[37]	LIU L,CAO P L,ZHANG L P,et al.Comparisons of adipogenesis- and lipid metabolism-related gene expression levels in muscle,adipose tissue and liver from Wagyu-cross and Holstein steers[J].PLoS One, 2021,16(2):e0247559.
[38]	TUVIA N,PIVOVAROVA-RAMICH O,MURAHOVSCHI V,et al.Insulin directly regulates the circadian clock in adipose tissue[J].Diabetes, 2021,70(9):1985-1999.
[39]	XIAO F,TANG C Y,TANG H N,et al.Long non-coding RNA 332443 inhibits preadipocyte differentiation by targeting Runx1 and p38-MAPK and ERK1/2-MAPK signaling pathways[J].Front Cell Dev Biol, 2021,9:663959.
[40]	ZHANG H Y,DENG T,GE S H,et al.Exosome circRNA secreted from adipocytes promotes the growth of hepatocellular carcinoma by targeting deubiquitination-related USP7[J].Oncogene, 2019,38(15):2844-2859.
[41]	LEFTEROVA M I,ZHANG Y,STEGER D J,et al.PPARγ and C/EBP factors orchestrate adipocyte biology via adjacent binding on a genome-wide scale[J].Genes Dev, 2008,22(21):2941-2952.
[42]	NIELSEN R,PEDERSEN T Å,HAGENBEEK D,et al.Genome-wide profiling of PPARγ:RXR and RNA polymerase II occupancy reveals temporal activation of distinct metabolic pathways and changes in RXR dimer composition during adipogenesis[J]. Genes Dev, 2008,22(21):2953-2967.
[43]	崔胜,王永,朱江江,等.过表达NR 4A1促进山羊皮下脂肪细胞分化[J].畜牧兽医学报,2021,52(5):1258-1266.CUI S,WANG Y,ZHU J J,et al.Overexpression of NR 4A1 promotes goat subcutaneous adipocytes differentiation[J].Acta Veterinaria et Zootechnica Sinica, 2021,52(5):1258-1266.(in Chinese)
[44]	JOO S T,HWANG Y H,FRANK D.Characteristics of Hanwoo cattle and health implications of consuming highly marbled Hanwoo beef[J].Meat Sci, 2017,132:45-51.
[45]	JUNG E Y,HWANG Y H,JOO S T.The relationship between chemical compositions,meat quality,and palatability of the 10 primal cuts from hanwoo steer[J].Korean J Food Sci Anim Resour, 2016,36(2):145-151.
[46]	HUANG J P,FENG X,ZHU R R,et al.Comparative transcriptome analysis reveals that PCK1 is a potential gene affecting IMF deposition in buffalo[J].BMC Genomics, 2020,21(1):710.

Overexpression of circNMT1 Promotes Adipogenic Differentiation of Buffalo Adipocytes

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References 46

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	TU Yun, ZENG Yanan, ZHANG Zhenghao, HONG Rui, WANG Zhen, WU Ping, ZHOU Zeyang, YE Yiru, DU Yanan, ZUO Fuyuan, ZHANG Gongwei. Genetic Structure and Runs of Homozygosity Analysis of Fuling Buffalo and Southwest Buffalo Breeds [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 1989-1998.
[2]	LIU Weiye, HUANG Xuewei. Research Progress of Non-coding RNA in Infectious Bursal Disease Virus Infection [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1488-1498.
[3]	YU Zhou, YANG Baigao, ZHANG Hang, XU Xi, ZHANG Peipei, FENG Xiaoyi, CAO Jianhua, NIU Yifan, DU Weihua, HAO Haisheng, ZHU Huabin, ABULIZI·Wusiman, ZHAO Xueming. Research Progress of Estrus Markers in Dairy Buffalo [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3623-3630.
[4]	TANG Yinmei, LI Qi, LI Haiyang, LIN Yaqiu, WANG Yong, XIANG Hua, HUANG Lian, ZHU Jiangjiang. Cloning of the Goat FATP2 Gene and Its Regulatory Effect on Lipid Deposition in Precursor Adipocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3642-3652.
[5]	SHAO Peng, TANG Yinmei, LIN Yaqiu, WANG Yong, XIANG Hua, HUANG Lian, ZHU Jiangjiang. Regulation Effect of PSMD9 on Lipid Deposition in Goat Precursor Adipocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3653-3663.
[6]	LIANG Rui, FAN Xiaorui, ZHANG Jinqiang, PANG Quanhai. Effects of Mouse Melanocyte Silencing and Overexpression of Pigment Epithelium-Derived Factor on Melanin Synthesis [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3916-3930.
[7]	LIU Linli, PENG Xuelan, LI Bo, CHENG Lefan, CIREN Lamu, ZHANG Enping. Effect of Overexpression of UCP3 Gene on the Differentiation of Sahu Sheep Preadipocytes [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3275-3285.
[8]	GUO Yixin, WANG Zhisheng, HU Rui, WANG Junmei, WANG Sen, SHI Liyuan, ZHANG Xiaohong, ZOU Huawei, ZUO Jiaxue, PENG Quanhui, XUE Bai, WANG Lizhi. Effect of Leucine on Browning of Subcutaneous Adipocytes in Yellow Cattle [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3286-3298.
[9]	WANG Wanjie, CHEN Nanzhu, ZOU Huiying, ZHOU Xinyi, HAO Haisheng, PANG Yunwei, ZHU Huabin, ZHAO Xueming, YU Dawei, DU Weihua. Effects of Histone Methyltransferase ASH1L Overexpression on Proliferation and Apoptosis of Bovine Cumulus Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3358-3368.
[10]	WANG Zhengrong, MA Xun, ZHANG Yanyan, SUN Yan, MENG Jimeng, BO Xinwen. Differential Expression Profile of CircRNA in Protoscolex, Hydatid Cyst Wall and Adult of Echinococcus granulosus [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3474-3489.
[11]	AN Zongqi, ZHAN Siyuan, LI Li, ZHANG Hongping. ceRNA-mediated Function of CircRNA on Critical Economic Traits in Animals [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(6): 2215-2222.
[12]	QUBI Wuqie, LI Yanyan, LI Xin, WANG Yong, WANG Youli, LIU Wei, ZHU Jiangjiang, LIN Yaqiu. APOA4 Gene Inhibits Intramuscular Adipocyte Differentiation in Goat [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 1927-1938.
[13]	QIN Xue, SHA Yiwen, YANG Menghao, CAI Rui, PANG Weijun. Advances in Regulation of Non-coding RNA on Mammalian Endometrial Receptivity and Decidualization [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1347-1358.
[14]	DAN Yixin, YANG Lu, XIANG Hua, ZHANG Huanrong, REN Yupeng, YANG Falong, HE Honghong, ZHU Jiangjiang. Effects of BIRC5 on the Cycle and Apoptosis of Goat Testis Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(4): 1511-1524.
[15]	MENG Qiuchi, LU Guangyu, CHEN Dingshuang, LIN Yaqiu, WANG Ruilong, ZHONG Chaosong, WANG Yong, LIU Wei, WANG Youli, LI Yanyan, LI Zhixiong. Goats GPR35 Gene Expression Characteristics Analysis and the Effect on Subcutaneous Fat Cell Differentiation [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(12): 4993-5007.