

Acta Veterinaria et Zootechnica Sinica ›› 2025, Vol. 56 ›› Issue (9): 4303-4314.doi: 10.11843/j.issn.0366-6964.2025.09.015
• Animal Genetics and Breeding • Previous Articles Next Articles
TIAN Jiao1(
), LONG Juyan1, CHEN Xia1, CEN Xiaoli1, NIU Xi1, HUANG Shihui2, WANG Jiafu1,*(
), RAN Xueqin2,*(
)
Received:2025-02-20
Online:2025-09-23
Published:2025-09-30
Contact:
WANG Jiafu, RAN Xueqin
E-mail:1037436332@qq.com;jfwang@gzu.edu.cn;xqran@gzu.edu.cn
CLC Number:
TIAN Jiao, LONG Juyan, CHEN Xia, CEN Xiaoli, NIU Xi, HUANG Shihui, WANG Jiafu, RAN Xueqin. Down-regulation of Gene Expression by SINE Insertion in the 3'UTR of the ENTPD1 Gene in the Xiang Pig[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(9): 4303-4314.
Table 1
Primers information"
| 引物名称 Primer name | 引物序列(5′→3′) Primer sequence | 产物长度/bp Length | 退火温度/℃ Annealing temperature | 用途 Application |
| ENTPD1-3′UTR-F | GAACTGAGAATCCTGAGTCCTGT | 571/274 | 58 | SV分型 |
| ENTPD1-3′UTR-R | CATAAGTTCCTTCACAAGCCTCT | |||
| qENTPD1-F | CTCCCACTCCACCTATGTCTTC | 160 | 60 | mRNA水平分析 |
| qENTPD1-R | AGTTTTCTTCTCACTCCAGCCA | |||
| qGAPDH-F | AGGTCGGAGTGAACGGATTTG | 118 | 60 | |
| qGAPDH-R | ACCATGTAGTGGAGGTCAATGAAG |
Table 2
Prediction of RBP binding sites in ENTPD1 3′UTR SINE"
| 序号 Number | RNA结合蛋白 RNA-binding protein | 结合位置 Binding position | 结合序列 Binding sequence | 数量 Number | 分数 Score |
| 1 | 富脯氨酸和谷氨酰胺的剪接因子(SFPQ) | 112~118 | GUGGUGU | 1 | 0.58 |
| 2 | Lin-28同系物A(LIN28A) | 113~119;159~165 | UGGUGUA | 2 | 0.98 |
| 3 | 多聚结合蛋白1(PCBP1) | 194~200 67~73 | CCUAGCC CCUGCCC | 1 1 | 0.72 0.56 |
| 5 | 脆性X信使核糖核蛋白1(FMR1) | 35~41 | CGACUAG | 1 | 0.83 |
| 6 | 富丝氨酸和精氨酸的剪接因子7(SRSF7) | 33~40 | UCCGACUA | 1 | 0.99 |
| 7 | 富丝氨酸和精氨酸的剪接因子9(SRSF9) | 14~20 | UGGCGCA | 1 | 0.99 |
| 8 | FUS RNA结合蛋白(FUS) | 114~117;160~163 | GGUG | 2 | 0.67 |
| 9 | FMR1常染色体同系物2(FXR2) | 126~132 | AGACGCG | 1 | 0.97 |
| 10 | 胰岛素样生长因子2 mRNA结合蛋白1(IGF2BP1) | 138~148 | GAUCCUGCGUU | 1 | 0.60 |
| 11 | 多聚嘧啶区结合蛋1(PTBP1) | 72~78 | CCUUGCU | 1 | 0.51 |
| 12 | Pumilio RNA结合家族成员1(PUM1) | 116~119;162~165 | UGUA | 2 | 0.67 |
Table 3
Prediction of miRNA binding sites in ENTPD1 3′UTR SINE"
| 序号 Number | miRNA名称 miRNA | 结合位置 Binding position | 序列(5′→3′) Sequence | 自由能/(kJ·mol-1) Free energy |
| 1 | ssc-miR-744 | 209~217 | AGCTCCGAT | -10.67 |
| 2 | ssc-miR-4331-3p | 209~215 | AGCTCCG | -11.61 |
| 3 | ssc-miR-214-5p | 195~203 | AGGCTGGCA | -10.52 |
| 4 | ssc-miR-9802-3p | 100~107 | GCCCTTGC | -11.57 |
| 5 | ssc-miR-326 | 258~266 | GCCCAAAGA | -11.06 |
| 6 | ssc-miR-330 | 258~266 | GCCCAAAGA | -12.02 |
| 7 | ssc-miR-885-3p | 128~135 | CGTTGCCG | -10.32 |
| 8 | ssc-miR-99b | 84~91 | TGCGGGTT | -10.75 |
| 9 | ssc-miR-652 | 78~86 | TGGGGTTGC | -10.23 |
| 10 | ssc-miR-9805-3p | 168~176 | GATCCTGCG | -10.87 |
| 11 | ssc-miR-7137-5p | 34~42 | GTTCCCGTC | -10.62 |
| 12 | ssc-miR-193a-5p | 254~262 | AAAGACCAA | -14.65 |
| 13 | ssc-miR-140-5p | 255~263 | AAGACCAAA | -11.56 |
Table 4
Population polymorphism analysis of ENTPD1 3′UTR SINE insertions/deletions"
| 群体 Population | 样本数/头 Sample | 基因型频率(%) Genotype frequency | 等位基因频率/% Allele frequency | HWE test P | 群体遗传特征 Population genetic characteristics | |||||||
| SINE+/+ | SINE+/- | SINE-/- | SINE+ | SINE- | 纯合度 Ho | 杂合度 He | 有效等位 基因数Ne | 多态信息 含量PIC | ||||
| 香猪 Xiang pig | 302 | 70 (23.18) | 124 (41.06) | 108 (35.76)A | 43.71 | 56.29A | 0.13 | 0.51 | 0.49 | 1.97 | 0.37 | |
| 大白猪 Large White pig | 170 | 54 (31.76) | 102 (60.00) | 14 (8.24)C | 61.76 | 38.24B | 0.03 | 0.53 | 0.47 | 1.90 | 0.36 | |
| 1 |
LI X Y , MOESTA A K , XIAO C , et al. Targeting CD39 in cancer reveals an extracellular ATP- and inflammasome-driven tumor immunity[J]. Cancer Discov, 2019, 9 (12): 1754- 1773.
doi: 10.1158/2159-8290.CD-19-0541 |
| 2 |
SCHADLICH I S , WINZER R , STABERNACK J , et al. The role of the ATP-adenosine axis in ischemic stroke[J]. Semin Immunopathol, 2023, 45 (3): 347- 365.
doi: 10.1007/s00281-023-00987-3 |
| 3 | WANG C , YI NQ , SN Z , et al. Progress on role of extracellular ATP and its metabolite adenosine in immunoregulation: Review[J]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi, 2020, 36 (12): 1134- 1140. |
| 4 |
GRASSI F , MARINO R . The P2X7 receptor in mucosal adaptive immunity[J]. Purinergic Signal, 2024, 20 (1): 9- 19.
doi: 10.1007/s11302-023-09939-w |
| 5 |
VUERICH M , ROBSON S C , LONGHI M S . Ectonucleotidases in intestinal and hepatic inflammation[J]. Front Immunol, 2019, 10, 507.
doi: 10.3389/fimmu.2019.00507 |
| 6 |
GUTKNECHT DA SILVA J L , PASSOS D F , CABRAL F L , et al. Istradefylline induces A2A/P2X7 crosstalk expression inducing pro-inflammatory signal, and reduces AKT/mTOR signaling in melanoma-bearing mice[J]. Med Oncol, 2023, 40 (6): 178.
doi: 10.1007/s12032-023-02033-6 |
| 7 |
PENG Z W , ROTHWEILER S , WEI G , et al. The ectonucleotidase ENTPD1/CD39 limits biliary injury and fibrosis in mouse models of sclerosing cholangitis[J]. Hepatol Commun, 2017, 1 (9): 957- 972.
doi: 10.1002/hep4.1084 |
| 8 |
LEE N T , SAVVIDOU I , SELAN C , et al. Development of endothelial-targeted CD39 as a therapy for ischemic stroke[J]. J Thromb Haemost, 2024, 22 (8): 2331- 2344.
doi: 10.1016/j.jtha.2024.04.023 |
| 9 |
ZENG J , NING Z , WANG Y , et al. Implications of CD39 in immune-related diseases[J]. Int Immunopharmacol, 2020, 89, 107055.
doi: 10.1016/j.intimp.2020.107055 |
| 10 |
SAVIO L E B , DE ANDRADE MELLO P , FIGLIUOLO V R , et al. CD39 limits P2X7 receptor inflammatory signaling a nd attenuates sepsis-induced liver injury[J]. J Hepatol, 2017, 67 (4): 716- 726.
doi: 10.1016/j.jhep.2017.05.021 |
| 11 |
MALONEY J P , BRANCHFORD B R , BRODSKY G L , et al. The ENTPD1 promoter polymorphism -860 A>G (rs3814159) is associated with increased gene transcription, protein expression, CD39/NTPDase1 enzymatic activity, and thromboembolism risk[J]. Faseb j, 2017, 31 (7): 2771- 2784.
doi: 10.1096/fj.201600344R |
| 12 |
ZHAO P , GU L , GAO Y , et al. Young SINEs in pig genomes impact gene regulation, genetic diversity, and complex traits[J]. Commun Biol, 2023, 6 (1): 894.
doi: 10.1038/s42003-023-05234-x |
| 13 |
TAM P L F , LEUNG D . The molecular impacts of retrotransposons in development and diseases[J]. Int J Mol Sci, 2023, 24 (22): 16418.
doi: 10.3390/ijms242216418 |
| 14 | ZHENG Y , CHEN C , WANG M , et al. SINE insertion in the pig carbonic anhydrase 5B (CA5B) gene is associated with changes in gene expression and phenotypic variation[J]. Animals (Basel), 2023, 13 (12): 1942. |
| 15 |
KRAMEROV D A , VASSETZKY N S . Origin and evolution of SINEs in eukaryotic genomes[J]. Heredity (Edinb), 2011, 107 (6): 487- 495.
doi: 10.1038/hdy.2011.43 |
| 16 |
SENFT A D , MACFARLAN T S . Transposable elements shape the evolution of mammalian development[J]. Nat Rev Genet, 2021, 22 (11): 691- 711.
doi: 10.1038/s41576-021-00385-1 |
| 17 |
CHOI J D , DEL PINTO L A , SUTTER N B . SINE retrotransposons import polyadenylation signals to 3'UTRs in dog (Canis familiaris)[J]. Mob DNA, 2025, 16 (1): 1.
doi: 10.1186/s13100-024-00338-5 |
| 18 |
WANG X , CHI C , HE J , et al. SINE insertion may act as a repressor to affect the expression of pig LEPROT and growth traits[J]. Genes (Basel), 2022, 13 (8): 1422.
doi: 10.3390/genes13081422 |
| 19 |
QI F , CHEN X , WANG J , et al. Genome-wide characterization of structure variations in the Xiang pig for genetic resistance to African swine fever[J]. Virulence, 2024, 15 (1): 2382762.
doi: 10.1080/21505594.2024.2382762 |
| 20 | 梁小芬, 冉雪琴, 牛熙, 等. 香猪CYP3A29基因3'-UTR结构变异及其与肌肉雄激素水平的关联分析[J]. 中国畜牧兽医, 2023, 50 (8): 3199- 3209. |
| LIANG X F , RAN X Q , NIU X , et al. Analysis of 3'-UTR Structural variations of CYP3A29 gene in Xiang pigs and its association with muscle androgen levels[J]. China Animal Husbandry & Veterinary Medicine, 2023, 50 (8): 3199- 3209. | |
| 21 |
PENG Z W , ROTHWEILER S , WEI G , et al. The ectonucleotidase ENTPD1/CD39 limits biliary injury and fibrosis in mouse models of sclerosing cholangitis[J]. Hepatol Commun, 2017, 1 (9): 957- 972.
doi: 10.1002/hep4.1084 |
| 22 |
SABATEL C , BUREAU F . The innate immune brakes of the lung[J]. Front Immunol, 2023, 14, 1111298.
doi: 10.3389/fimmu.2023.1111298 |
| 23 |
RYANTO G R T , SURAYA R , NAGANO T . The importance of lung innate immunity during health and disease[J]. Pathogens, 2025, 14 (1): 91.
doi: 10.3390/pathogens14010091 |
| 24 |
WANG L , LI Y J , YANG X , et al. Purinergic signaling: a potential therapeutic target for ischemic stroke[J]. Purinergic Signal, 2023, 19 (1): 173- 183.
doi: 10.1007/s11302-022-09905-y |
| 25 |
ROBSON S C , WU Y , SUN X , et al. Ectonucleotidases of CD39 family modulate vascular inflammation and thrombosis in transplantation[J]. Semin Thromb Hemost, 2005, 31 (2): 217- 233.
doi: 10.1055/s-2005-869527 |
| 26 |
ORTIZ M A , DIAZ-TORNÉ C , DE AGUSTIN J J , et al. Altered CD39 and CD73 expression in rheumatoid arthritis: Implications for disease activity and treatment response[J]. Biomolecules, 2023, 14 (1): 1.
doi: 10.3390/biom14010001 |
| 27 | 李默晗. CD39通过下调PAI-1表达与活性保护缺血性卒中脑组织[D]. 长沙: 中南大学, 2022. |
| LI M H. CD39 protects the brain tissue of ischemic stroke by down-regulating the expression and activity of PAI-1[D]. Changsha: Zhongnan University, 2022. (in Chinese) | |
| 28 | 夏国庆. ATP通过P2X4和CD39调控酒精相关性脂肪性肝炎机制研究[D]. 合肥: 安徽医科大学, 2022. |
| XIA G Q. Research on the Mechanism of ATP regulating Alcoholic Steatohepatitis through P2X4 and CD39[D]. Hefie: Anhui Medical University, 2022. (in Chinese) | |
| 29 | 杜鹏. CD39-腺苷-A2B信号通路在低温携氧机械灌注改善DCD肝脏缺血再灌注损伤中的作用及机制研究[D]. 南昌: 南昌大学, 2022. |
| DU P. Role and mechanism of CD39-adenosine-A2B signaling pathway in hypothermic oxygen-carrying mechanical perfusion to ameliorate liver ischemia-reperfusion injury in DCD[D]. Nanchang: Nanchang University, 2022. (in Chinese) | |
| 30 |
LIU J J , RAN X Q , LI S , et al. Polymorphism in the first intron of follicle stimulating hormone beta gene in three Chinese pig breeds and two European pig breeds[J]. Anim Reprod Sci, 2009, 111 (2-4): 369- 375.
doi: 10.1016/j.anireprosci.2008.03.004 |
| 31 |
MAGOTRA A , NASKAR S , DAS B , et al. A comparative study of SINE insertion together with a mutation in the first intron of follicle stimulating hormone beta gene in indigenous pigs of India[J]. Mol Biol Rep, 2015, 42 (2): 465- 470.
doi: 10.1007/s11033-014-3789-y |
| 32 | 黄月丽, 冉雪琴, 牛熙, 等. 香猪CHD3基因3'-侧翼区264 bp结构变异的研究[J]. 中国畜牧兽医, 2022, 49 (8): 3026- 3035. |
| HUANG Y L , RAN X Q , NIU X , et al. A study of 264 bp structural variation in the 3'-flanking region of the CHD3 gene of the balsam pig[J]. China Animal Husbandry & Veterinary Medicine, 2022, 49 (8): 3026- 3035. | |
| 33 |
ZHAO P , GU L , GAO Y , et al. Young SINEs in pig genomes impact gene regulation, genetic diversity, and complex traits[J]. Commun Biol, 2023, 6 (1): 894.
doi: 10.1038/s42003-023-05234-x |
| 34 | TUCKER J M , GLAUNSINGER B A . Host noncoding retrotransposons induced by DNA viruses: a SINE of infection?[J]. J Virol, 2017, 91 (23): e00982- 17. |
| 35 |
KARIJOLICH J , ABERNATHY E , GLAUNSINGER B A . Infection-induced retrotransposon-derived noncoding RNAs enhance herpesviral gene expression via the NF-κB pathway[J]. PLoS Pathog, 2015, 11 (11): e1005260.
doi: 10.1371/journal.ppat.1005260 |
| 36 |
HWANG H , CHANG H R , BAEK D . Determinants of functional microRNA targeting[J]. Mol Cells, 2023, 46 (1): 21- 32.
doi: 10.14348/molcells.2023.2157 |
| 37 |
XI E , BAI J , ZHANG K , et al. Genomic variants disrupt miRNA-mRNA regulation[J]. Chem Biodivers, 2022, 19 (10): e202200623.
doi: 10.1002/cbdv.202200623 |
| 38 |
李河林, 蒋玉芬, 程娜, 等. 筛选的差异表达microRNAs对猪卵母细胞Npm2基因的表达调控及作用研究[J]. 畜牧兽医学报, 2024, 55 (11): 5035- 5049.
doi: 10.11843/j.issn.0366-6964.2024.11.021 |
|
LI H L , JIANG Y F , CHENG N , et al. Regulation of Npm2 gene expression and role of screened differentially expressed microRNAs in porcine oocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55 (11): 5035- 5049.
doi: 10.11843/j.issn.0366-6964.2024.11.021 |
|
| 39 |
KOH D , BIN JEON H , OH C , et al. RNA-binding proteins in cellular senescence[J]. Mech Ageing Dev, 2023, 214, 111853.
doi: 10.1016/j.mad.2023.111853 |
| 40 |
余祖华, 高梦茹, 齐志颖, 等. RNA结合蛋白ELAVL1的功能及其调控病毒复制的研究进展[J]. 畜牧兽医学报, 2024, 55 (5): 1914- 1925.
doi: 10.11843/j.issn.0366-6964.2024.05.010 |
|
YU Z H , GAO M R , QI Z Y , et al. Advances in the function of the RNA-binding protein ELAVL1 and its regulation of viral replication[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55 (5): 1914- 1925.
doi: 10.11843/j.issn.0366-6964.2024.05.010 |
|
| 41 |
AMEIS D , LIU F , KIRBY E , et al. The RNA-binding protein Quaking regulates multiciliated and basal cell abundance in the developing lung[J]. Am J Physiol Lung Cell Mol Physiol, 2021, 320 (4): L557- L567.
doi: 10.1152/ajplung.00481.2019 |
| [1] | GUI Ruohong, CAO Hongzhan, LIU Songzan, LIU Jixiang, ZHAO Jialong, LU Chunlian. Effects of Different Dietary Metabolizable Energy and Standard Ileal Digestible Lysine Levels on Performance of High-Producing Lactating Shenxian Sows [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(9): 4472-4490. |
| [2] | ZONG Yunhe, YANG Yuze, SUN Yanyan, CHEN Jilan, LI Yunlei. Research Advances in the Investigation of the Protective Role of Lysine Acetylation in Chicken Semen Cryopreservation and Its Mechanism [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(7): 3071-3079. |
| [3] | CHEN Zhihua, WANG Qi, ZHANG Jin, YANG Liandi, YANG Tianqing, WANG Jing, LONG Dingbiao, HUANG Jinxiu, HUANG Wenming. Effects of Dietary Net Energy and Lysine Levels on Reproductive Performance, Serum Hormone, Lactation Performance and Fecal Flora Diversity of Rongchang Sows in Late Pregnancy [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(6): 2801-2815. |
| [4] | LIU Xinzi, ZHAO Haiyuan, JU Ning, CHEN Ying, WANG Zi, MENG Weijing, LI Jiaxuan, JIANG Yanping, CUI Wen, WANG Xiaona. Effect of Constitutive Promoter Pldh on Exogenous Gene Expression in Lactic Acid Bacteria Expression System [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(6): 2937-2947. |
| [5] | ZHOU Rui, WU De, CHE Lianqiang, LIN Yan, FENG Bin, FANG Zhengfeng. Advances of N6-Adenosine Methylation Regulating Adipogenesis [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(5): 1995-2003. |
| [6] | MA Yingtian, JIANG Luyao, LI Zengkai, QIN Jianping, ZHAO Jianhua, HE Yufang, SONG Yuxuan, ZHANG Lei. Effect of Cyanidin-3-rutinoside on Cryopreservation of Semen of Dairy Sheep [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(4): 1768-1778. |
| [7] | ZHOU Xianshan, HUANG Shihui, NIU Xi, RAN Xueqin, WANG Jiafu. Differential Expression Study of Structural Variation in the Ubiquitin Ligase 2 Gene of Xiang Pigs with Wrinkled Skin [J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(1): 136-146. |
| [8] | LONG Tanghui, ZHAN Yanbo, LIAO Guanxiang, CHEN Xinfeng, ZHANG Jian, LI Yanjiao, OUYANG Kehui, QIU Qinghua. Effects of Dietary Lysine Supplementation on Fecal Fermentation Parameters and Microbial Flora Structure of Beef Cattle [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(5): 2042-2049. |
| [9] | YANG Yang, YU Qian, LIU Yucheng, YANG Hua, ZHAO Zhuo, WANG Limin, ZHOU Ping, YANG Qingyong, DAI Rong. Identification and Tissue Expression Analysis of the Sheep MYL Gene Family [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1551-1564. |
| [10] | KANG Jia, DUAN Xiangru, YIN Xuejiao, YANG Ruochen, LI Taichun, SHAN Xinyu, CHEN Meijing, ZHANG Yingjie, LIU Yueqin. Effects of Cysteine and Methionine on Secondary Hair Follicle Growth and Hair Dermal Papilla Cell Proliferation in vitro in Cashmere Goats [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 515-527. |
| [11] | YAN Chao, LIU Yonggang, XIE Hao, PENG Cuiting, ZHANG Caiyong, ZHAO Yulan, QI Lin, CHEN Zhilong, TANG Zhonglin. Detection of Gene Expression in Trace Cells of Early Porcine Embryo by Pre-amplified Quantitative PCR [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5567-5574. |
| [12] | Yongqing LIU, Gang ZHANG, Yanling XIONG, Zhongxin SUN, Fan GAO, Ting LIU, Hui LI. Effects of Heat Stress on Duodenal Mucosal Structure, HIF-1 and Its Related Protein Expression in Congjiang Xiang Pigs [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(10): 4690-4699. |
| [13] | YUAN Wei, BI Huan, ZHANG Yudan, ZHANG Yiyu, GU Xiaolong, YANG Hongwen, CHEN Wei. Deciphering Genome-wide Selection Signals Reveals Genetic Differences between Jianbai and Congjiang Xiang Pigs [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3631-3641. |
| [14] | SUI Yuzhen, CHEN Guizhen, ZUO Shoujun, WANG Haidong, SONG Pengtao, LI Liangliang, ZHANG Longxian, DONG Haiju, LIU Fang. Analysis on Infection Status of Clonorchis sinensis and Other Intestinal Parasites in Pet Cats in Henan Province [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(7): 3022-3030. |
| [15] | LI Yujuan, ZHANG Yuanming, ZHANG Beiyu, LI Fuchang, LIU Lei. Effects of Dietary Lysine Supplementation on Hair Production Performance and Hair Follicle Development of Angora Rabbits [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2013-2019. |
| Viewed | ||||||
|
Full text |
|
|||||
|
Abstract |
|
|||||