猪miR-1343生物信息学分析及其在卵泡颗粒细胞中的表达研究

doi:10.16431/j.cnki.1671-7236.2025.08.022

Abstract

Abstract: 【Objective】 The purpose of this paper was to investigate the regulatory mechanism of miR-1343 in porcine ovarian granulosa cells.The target genes of miR-1343 were predicted and analyzed by the bioinformatics method，and the effects of reproductive hormone on miR-1343 expression were explored in porcine ovarian granulosa cells. 【Method】 Mature sequences of miR-1343 were downloaded from the miRbase online database for sequence conservative analysis.Porcine ovarian granulosa cells were isolated and cultured,which were seeded into six-well plates.At 70%-80% cell confluence,the granulosa cells were treated with 0 IU/mL (control group) and 20 IU/mL (experimental group) follicle-stimulating hormone (FSH),with 3 replicates per group,24 h after addition,the cells were harvested,and the expression of miR-1343 was detected by Real-time quantitative PCR.The PROMO online software was utilized to predict the transcription factor binding sites for miR-1343 in pigs.The target genes of miR-1343 were predicted using TargetScan,miRDB,and Starbase database.GO function and KEGG pathway enrichment analysis were performed for the target genes using DAVID online database.The protein-protein interaction (PPI) of target genes was analyzed using STRING database,and regulatory network was constructed using Cytoscape software. 【Result】 The mature sequences of miR-1343 were highly conserved among multiple species.The relative expression of miR-1343 in experimental group was extremely significantly down-regulated compared with the control group (P＜0.01).There were many transcription factor binding sites such as Sp1,SMAD3,SMAD4,and FOXO4 in the promoter region of miR-1343.A total of 278 common target genes were predicted using different databases.GO function annotation analysis results revealed that target genes of miR-1343 were mainly enriched in phosphorylation,positive regulation of transcription by RNA polymerase Ⅱ,protein phosphorylation,and positive regulation of DNA-templated transcription for biological process,nucleus,cytoplasm,cytosol,and nucleoplasm for cellular component,and ATP binding,protein homodimerization activity,protein serine/threonine kinase activity and chromatin binding for molecular function.KEGG analysis results revealed that target genes of miR-1343 were significantly enriched in the Hippo signaling pathway,MAPK signaling pathway,and thyroid hormone signaling pathway,which were related to ovarian function.PPI analysis of miR-1343 target genes indicated that SYNJ1,PLCB1,and GSK3B had more targeted relationships with other proteins. 【Conclusion】 The expression of miR-1343 was influenced by FSH stimulation in granulosa cells.miR-1343 might participate in the development of porcine ovarian granulosa cells by regulating the expression of target genes such as PLCB1 and GSK3B.The results provided a reference for further exploring the regulatory mechanism of miR-1343 in porcine ovarian granulosa cells.

Key words: porcine ovarian granulosa cells; miR-1343; target gene; bioinformatics

CLC Number:

S813.3

HU Huiyan, SHAO Liwei, LIU Xiaohui, YUAN Ying, PI Mingwei. Bioinformatics Analysis of miR-1343 and Its Expression in Porcine Ovarian Granulosa Cells[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(8): 3734-3743.

References

[1] VAISHNAV S,CHAUHAN A,AJAY A,et al.Allelic to genome wide perspectives of swine genetic variation to litter size and its component traits[J].Molecular Biology Reports,2023,50(4):3705-3721.
[2] JIANG Y,HE Y,PAN X,et al.Advances in oocyte maturation in vivo and in vitro in mammals[J].International Journal of Molecular Sciences,2023,24(10):9059.
[3] MATSUDA F,INOUE N,MANABE N,et al.Follicular growth and atresia in mammalian ovaries:Regulation by survival and death of granulosa cells[J].Journal of Reproduction and Development,2012,58(1):44-50.
[4] YU Y S,SUI H S,HAN Z B,et al.Apoptosis in granulosa cells during follicular atresia:Relationship with steroids and insulin-like growth factors[J].Cell Research,2004,14(4):341-346.
[5] ZHANG H,LIU K.Cellular and molecular regulation of the activation of mammalian primordial follicles:Somatic cells initiate follicle activation in adulthood[J]. Human Reproduction Update,2015,21(6):779-786.
[6] SONTAKKE S D,MOHAMMED B T,MCNEILLY A S,et al.Characterization of microRNAs differentially expressed during bovine follicle development[J].Reproduction,2014,148(3):271-283.
[7] ZHONG Y,LI L,CHEN Z,et al.miR143 inhibits steroidogenesis and induces apoptosis repressed by H3K27 me3 in granulosa cells[J].Frontiers in Cell and Developmental Biology,2020,8:565261.
[8] DU X,WANG L,LI Q,et al.miR-130a/TGF-β1 axis is involved in sow fertility by controlling granulosa cell apoptosis[J].Theriogenology,2020,157:407-417.
[9] JIANG L,HUANG H,QIAN Y,et al.miR-130b regulates gap junctional intercellular communication through connexin 43 in granulosa cells from patients with polycystic ovary syndrome[J].Molecular Human Reproduction,2020,26(8):576-584.
[10] PERSSON H,KVIST A,REGO N,et al.Identification of new microRNAs in paired normal and tumor breast tissue suggests a dual role for the ERBB2/Her2 gene[J].Cancer Research,2011,71(1):78-86.
[11] TIAN Y,ZHANG M Y,LI N,et al.Zearalenone exposure triggered porcine granulosa cells apoptosis via microRNAs-mediated focal adhesion pathway[J].Toxicology Letters,2020,330(5):80-89.
[12] ZHANG R,ZHOU Z,WANG P,et al.The SLC19A1-AS/miR-1343/WNT11 axis is a novel positive regulatory ceRNA network governing goat granulosa cell proliferation[J].International Journal of Biological Macromolecules,2024,264:130658.
[13] PASQUARIELLO R,MANZONI E F M,FIANDANESE N,et al.Implications of miRNA expression pattern in bovine oocytes and follicular fluids for developmental competence[J].Theriogenology,2020,145:77-85.
[14] WANG P,LIU Z,ZHANG X,et al.Integrated analysis of lncRNA,miRNA and mRNA expression profiles reveals regulatory pathways associated with pig testis function[J].Genomics,2024,116(2):110819.
[15] HU H,JIA Q,XI J,et al.Integrated analysis of lncRNA,miRNA and mRNA reveals novel insights into the fertility regulation of Large White sows[J].BMC Genomics,2020,21(1):636.
[16] YIN L,WANG W,WEI H,et al.Localization and expression of CTRP6 in ovary and its regulation by FSH in porcine granulosa cells[J].Theriogenology,2019,127:56-65.
[17] SELBACH M,SCHWANHAUSSER B,THIERFELDER N,et al.Widespread changes in protein synthesis induced by microRNAs[J].Nature,2008,455(7209):58-63.
[18] GUO Y,DING X,DAI C,et al.miR13433p inhibits autophagy by directly targeting ATG7 in multiple myeloma cells[J].Biomedical Reports,2024,21(6):185.
[19] ZHOU Y,HUANG T,ZHANG J,et al.TEAD1/4 exerts oncogenic role and is negatively regulated by miR-4269 in gastric tumorigenesis[J].Oncogene,2017,36(47):6518-6530.
[20] ZHANG Y,WANG X,LIU W,et al.CircGLIS3 promotes gastric cancer progression by regulating the miR-1343-3p/PGK1 pathway and inhibiting vimentin phosphorylation[J].Journal of Translational Medicine,2024,22(1):251.
[21] STOLZENBURG L R,WACHTEL S,DANG H,et al.miR-1343 attenuates pathways of fibrosis by targeting the TGF-β receptors[J].Biochemical Journal,2016,473(3):245-256.
[22] XIE Y,CAO H,ZHANG Z,et al.Molecular network of miR-1343 regulates the pluripotency of porcine pluripotent stem cells via repressing OTX2 expression[J].RNA Biology,2019,16(1):82-92.
[23] QIN W,JIANG J,WU J,et al.Exosomal ssc-miR-1343 targets FAM131C to regulate Porcine epidemic diarrhea virus infection in pigs[J].Veterinary Research,2024,55(1):91.
[24] ZHOU J,LIU J,PAN Z,et al.The let-7g microRNA promotes follicular granulosa cell apoptosis by targeting transforming growth factor-β type 1 receptor[J].Molecular and Cellular Endocrinology,2015,409:103-112.
[25] HU H,FU Y,ZHOU B,et al.Long non-coding RNA TCONS_00814106 regulates porcine granulosa cell proliferation and apoptosis by sponging miR-1343[J].Molecular and Cellular Endocrinology,2021,520:111064.
[26] LI Q,DU X,WANG L,et al.TGF-β1 controls porcine granulosa cell states:A miRNA-mRNA network view[J].Theriogenology,2021,160:50-60.
[27] 林冬静,何倩丽.SP1介导的信号转导通路对恶性肿瘤形成的作用[J].中国组织化学与细胞化学杂志,2016,25(1):91-95. LIN D J,HE Q L.Role of SP1-mediated signal transduction pathways in cancer formation[J].Chinese Journal of Histochemistry and Cytochemistry,2016,25(1):91-95.(in Chinese)
[28] YANG L,DU X,LIU L,et al.miR-1306 mediates the feedback regulation of the TGF-β/SMAD signaling pathway in granulosa cells[J].Cells,2019,8(4):298.
[29] ZHANG H Y,WANG W,HE Y,et al.Effects of silencing smad4 gene by small interfering RNA on apoptosis of porcine granulosa cells[J].Chinese Journal of Cell Biology,2010,32(4):596-600.
[30] DU X,ZHANG L,LI X,et al.TGF-β signaling controls FSHR signaling-reduced ovarian granulosa cell apoptosis through the SMAD4/miR-143 axis[J].Cell Death & Disease,2016,7(11):e2476.
[31] MA M,ZHANG J,GAO X,et al.miR-361-5p mediates SMAD4 to promote porcine granulosa cell apoptosis through VEGFA[J].Biomolecules,2020,10(9):1281.
[32] ZHANG J Q,GAO B W,GUO H X,et al.miR-181a promotes porcine granulosa cell apoptosis by targeting TGFBR1 via the activin signaling pathway[J].Molecular and Cellular Endocrinology,2020,499:110603.
[33] CLARK K L,GEORGE J W,PRZYGRODZKA E,et al.Hippo signaling in the ovary:Emerging roles in development,fertility,and disease[J]. Endocrine Reviews,2022,43(6):1074-1096.
[34] CASTILHO A C,NOGUEIRA M F,FONTES P K,et al.Ovarian superstimulation using FSH combined with equine chorionic gonadotropin (eCG) upregulates mRNA-encoding proteins involved with LH receptor intracellular signaling in granulosa cells from Nelore cows[J].Theriogenology,2014,82(9):1199-1205.
[35] CHEN H L,CHENG J Y,YANG Y F,et al.Phospholipase C inhibits apoptosis of porcine oocytes cultured in vitro[J].Journal of Cellular Biochemistry,2020,121(7):3547-3559.
[36] SPATE L D,BROWN A N,REDEL B K,et al.Dickkopf-related protein 1 inhibits the Wnt signaling pathway and improves pig oocyte maturation[J].PLoS One,2014,9(4):e95114.
[37] BOTHUM A M,WOODS D C.Dynamics of Wnt signaling components in the human ovary from development to adulthood[J].Histochemistry and Cell Biology,2019,151(2):115-123.
[38] ISMAIL A B,NAJI M’S,NEBIH I·,et al.The expression profile of Wnt/β-catanin signalling genes in human oocytes obtained from polycystic ovarian syndrome (PCOS) patients[J].Zygote,2022,30(4):536-542.
[39] KAWAMURA K,CHENG Y,SUZUKI N,et al.Hippo signaling disruption and Akt stimulation of ovarian follicles for infertility treatment[J].Proceedings of the National Academy of Sciences of the United States of America,2013,110(43):17474-17479.
[40] WU H,LIU Q,YANG N,et al.Polystyrene-microplastics and DEHP co-exposure induced DNA damage,cell cycle arrest and necroptosis of ovarian granulosa cells in mice by promoting ROS production[J].Science of the Total Environment,2023,871:161962.

Bioinformatics Analysis of miR-1343 and Its Expression in Porcine Ovarian Granulosa Cells

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	PING Yuyu, HUANG Xuan, CAI Qingqing, WANG Qiangzhou, WANG Jiaxing, BAI Hao, CHEN Shihao, CHANG Guobin. Prediction of Biological Function of gga-miR-1574-5p and Verification of Its Targeting Relationship with G3BP2 Gene [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 2981-2991.
[2]	ZHENG Hao, LIU Mengyun, WANG Mingyu, DONG Xia, SHI Liangyu, YU Bo, YANG Yu, ZHOU Ao, CHEN Xing. Cloning, Bioinformatics and Tissue Expression Analysis of ARHGDIB Genes in Ducks [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 3004-3015.
[3]	LI Zhiquan, FU Sijing, XU Shuping, GAO Mengruo, YANG Taotao, ZHANG Zhibang, LI Kai, LI Pengcheng. Cloning, Bioinformatics Analysis and Tissue Expression of HSPA5 Gene in Wuyi Black Pig [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 3214-3224.
[4]	CHEN Xuke, PU Na, ZHANG Yuxia, ZHAO Wenqing, ZHAO Jiaxin, HUANG Juncheng, WEI Linying, ZHANG Yanyan, SUN Yan, BO Xinwen, WANG Zhengrong. Bioinformatics Analysis of Methionine Aminopeptidase Ⅱ of Echinococcus granulosus and Its Differential Expression in Different Stages of the Worm [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(7): 3286-3296.
[5]	YANG Quan, LI Xiao, YAN Zunqiang, WANG Pengfei, HUANG Xiaoyu, GAO Xiaoli, YANG Qiaoli, GUN Shuangbao, YANG Jiaojiao. Cloning,Bioinformatics Analysis and Tissue Expression of CXCL12 Gene in Hezuo Pigs [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2482-2493.
[6]	GUAN Song, SHI Liguang, LIN Yu, JIANG Jianxiao, WU Hongzhi, PENG Weiqi. Functional Prediction of GDF9 Gene nsSNP and Its Correlation Analysis with Litter Size in Hainan Black Goats [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(5): 2166-2176.
[7]	HU Huihui, FU Panpan, LI Jie, YAN Zunqiang, GAO Xiaoli, YANG Jiaojiao, HUANG Xiaoyu. Cloning,Identification and Tissue Expression Analysis of TRIF Gene CDS Region in Hezuo Pigs [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(4): 1478-1487.
[8]	LIU Xiaotong, GE Bingjie, QIU Qian, LIU Xinman, YU Minghong, ZHANG Xuemei. Application of Bioinformatics in Toxicity Mechanism of Aflatoxin B₁ [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(4): 1639-1650.
[9]	ZHANG Ying, CHEN Guangzhen, LIU Mei, LI Hongxia, YU Yan, JIA Xuebo, LI Wenqian, FAN Jianhua, MA Zhiyu. Cloning,Bioinformatics and Tissue Expression Analysis of NMB Gene in Mulard Duck [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(4): 1705-1715.
[10]	MIAO Shukui, MI Xiaoyun, WEI Jie, WEI Yurong, HAILIQIEMU·Maimaitiyiming. Cloning and Bioinformatics Analysis of VP7 Gene of the Genotype G8 Bovine Rotavirus [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(4): 1784-1795.
[11]	Siriguleng, YU Wen, JIANG Xiaowei, LI Ziyi, JIN Junjian, BAI Haoyu. The Expression of miR-144-5p in Plasma Exosomes of Different Body Types of Bactrian Camels and the Verification of Target Gene [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(3): 1022-1032.
[12]	AI Xiaonan, CHENG Lifen, BAI Pu, CHEN Zhenghao, XUE Lina, ZHOU Shenghua. Cloning and Bioinformatics Analysis of ACSL1 Gene and Its Expression Pattern During Adipocyte Differentiation in Lingqiu Qingbei Goats [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(2): 499-511.
[13]	LEI Zhigang, PAN Hong, ZHANG Dandan, LIU Quanhui, SUN Zhe, DENG Shan, HUANG Ben. Cloning, Bioinformatics Analysis and Eukaryotic Expression Vector Construction of MAPK8 Gene in Mashan Black Goats [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(2): 534-544.
[14]	ZHAO Wenqing, BO Xinwen, PU Na, ZHANG Yuxia, CHEN Xuke, ZHANG Yanyan, SUN Yan, QI Meng, WANG Zhengrong. Bioinformatics Analysis and Prokaryotic Expression of EgBAL Protein of Echinococcus granulosus [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(2): 801-810.
[15]	TIAN Huihui, WANG Bingxin, ZHU Zhaoyan, YU Yange, DING Mengxia, TIAN Yadong, JIANG Ruirui, SUN Guirong, HAN Ruili, KANG Xiangtao, YAN Fengbin, GUO Yujie. Cloning,Bioinformatics Analysis of HSPA8 Gene and Its Expression Pattern in Thymus in Chickens [J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(1): 39-51.