转录组分析探讨揭示羊肚菌多糖ME-X抗S180肿瘤的分子机制

doi:10.16036/j.issn.1000-2650.202110086

摘要/Abstract

摘要： 【目的】为探究羊肚菌多糖（ME-X）对S180肿瘤细胞抑制作用及其基因表达的影响,探索ME-X抑制小鼠S180肿瘤表型下潜在的作用分子及主要信号通路。【方法】以S180肿瘤小鼠为模型,先进行ME-X体内抑制小鼠S180肿瘤试验,再结合RNA-Seq测序技术对体内试验组中小鼠的S180肿瘤细胞进行测序并对得到的数据进行生物信息学分析。【结果】羊肚菌多糖（ME-X）和甘露聚糖肽（mannatide）一样能够极显著（P<0.01）抑制小鼠S180肿瘤的生长,其抑瘤率可达53.81%。差异表达基因（differential expressed genes,DEGs）分析结果显示羊肚菌多糖抑制S180肿瘤的关键基因主要富集在PI3K-AKT信号通路中,该通路中的成纤维细胞生长因子（Fgf10）、成纤维细胞生长因子受体（Fgfr2）、细胞因子受体（Prlr、Ghr、I12rb）、磷酸烯醇式丙酮酸羧激酶（Pck1）等多个基因显著上调。【结论】羊肚菌多糖（ME-X）可能通过PI3K-AKT信号通路调控S180肿瘤细胞代谢、迁移、细胞周期等进程达到抑制S180肿瘤的目的,为羊肚菌多糖抑制肿瘤分子机制研究提供可参考的理论依据。

关键词: 羊肚菌多糖, S180肿瘤, RNA-Seq, 差异表达基因, PI3K-AKT信号通路

Abstract: 【Objective】 This study aims to explore the antitumor activity of a polysaccharides from Morchella esculenta （named ME-X） and its impact on gene expression, observing the potential effect molecules and main signaling pathways under ME-X, which had a phenotype of suppressing the growth of S180 tumor in mice. 【Method】 In this study, the S180 tumor bearing mice were used as a model and antitumor experiments in mice were performed at first, then RNA-Seq technology was combined to sequence the S180 tumor cells from those mice in the experimental groups with bioinformatics analysis of the obtained data. 【Result】 ME-X, like Mannatide, which could significantly （P<0.01） inhibit the growth of S180 tumor in mice, and the rate of inhibition reached 53.81%. Differential expression genes （DEGs） analysis suggested that the key genes mostly enriched in the PI3K-AKT signaling pathway including many significantly up-regulated genes, such as fibroblast growth factor 10 （Fgf10）, fibroblast growth factor receptor 2 （Fgfr2）, cytokine receptors （Prlr, Ghr, I12rb） and phosphoenolpyruvate carboxykinase （Pck1）. 【Conclusion】 It was suggested that ME-X could regulate the metabolism, migration and cell cycle of S180 tumor cells through PI3K-AKT signaling pathway to inhibit S180 tumor.

Key words: ME-X, S180 tumor, RNA-Seq, differential expressed genes （DEGs）, PI3K-AKT signaling pathway

中图分类号:

Q591

鲁艳, 黄瑶, 叶姿妤, 周丽倩, 刘欣岚, 丁祥, 侯怡铃. 转录组分析探讨揭示羊肚菌多糖ME-X抗S180肿瘤的分子机制[J]. 四川农业大学学报, 2022, 40(4): 519-528.

LU Yan, HUANG Yao, YE Ziyu, ZHOU Liqian, LIU Xinlan, DING Xiang, HOU Yiling. Transcriptomes Analysis Revealed the Molecular Mechanism about the Anti-S180 Tumor Activity of the Polysaccharide ME-X from Morchella esculenta[J]. Journal of Sichuan Agricultural University, 2022, 40(4): 519-528.

参考文献

[1] DWEK R A.Glycobiology: toward understanding the function of sugars[J]. Chemical Reviews, 1996, 96(2): 683-720.
[2] YU Y, SHEN M Y, SONG Q, et al.Biological activities and pharmaceutical applications of polysaccharide from natural resources: a review[J]. Carbohydrate Polymers, 2018, 183: 91-101.
[3] CAI Z, LI W, WANG H T, et al.Anti-tumor and immunomodulating activities of a polysaccharide from the root of Sanguisorba officinalis L[J]. International Journal of Biological Macromolecules, 2012, 51(4): 484-488.
[4] ZONG S, LI J L, YE Z Y, et al.Lachnum polysaccharide suppresses S180 sarcoma by boosting anti-tumor immune responses and skewing tumor-associated macrophages toward M1 phenotype[J]. International Journal of Biological Macromolecules, 2020, 144: 1022-1033.
[5] 陈真,郭青龙,钱之玉,等. 海洋真菌多糖YCP对荷瘤小鼠的抗肿瘤作用[J]. 食品与生物技术学报, 2005, 24(5): 1673-1689.
[6] DUNCAN C J G, PUGH N, PASCO D S, et al. Isolation of a galactomannan that enhances macrophage activation from the edible fungus Morchella esculenta[J]. Journal of Agricultural and Food Chemistry, 2002, 50(20): 5683-5685.
[7] HU M L, CHEN Y, WANG C, et al.Induction of apoptosis in HepG2 cells by polysaccharide MEP-II from the fermentation broth of Morchella esculenta[J]. Biotechnology Letters, 2013, 35(1): 1-10.
[8] LIU C, SUN Y H, MAO Q, et al.Characteristics and antitumor activity of Morchella esculenta polysaccharide extracted by pulsed electric field[J]. International Journal of Molecular Sciences, 2016, 17(6): 986.
[9] 黄瑶, 蒋琳, 刘影, 等. 羊肚菌多糖提取、分离纯化及免疫调节活性[J]. 生物加工过程, 2018, 16(6): 35-41.
[10] 丁祥, 侯怡铃, 黄瑶. 一种羊肚菌多糖及其制备方法和应用[P].2021-9-10, 中国, ZL201910559463.4
[11] LIU Y H, ZHAO Y, TANG R, et al.Effect of TFAM on ATP content in tachypacing primary cultured cardiomyocytes and atrial fibrillation patients[J]. Molecular Medicine Reports, 2020, 22(6): 5105-5112.
[12] ACÍN-PÉREZ R, BAYONA-BAFALUY M P, FERNÁNDEZ-SILVA P, et al. Respiratory complex Ⅲ is required to maintain complex I in mammalian mitochondria[J]. Molecular Cell, 2004, 13(6): 805-815.
[13] SINGH R K, SAINI S K, PRAKASAM G, et al.Role of ectopically expressed mtDNA encoded cytochrome c oxidase subunit I (MT-COI) in tumorigenesis[J]. Mitochondrion, 2019, 49: 56-65.
[14] DUDKINA N V, KOURIL R, PETERS K, et al.Structure and function of mitochondrial supercomplexes[J]. Biochimica et Biophysica Acta, 2010, 1797(6-7): 664-670.
[15] MICK D U, FOX T D, REHLING P.Inventory control: cytochrome c oxidase assembly regulates mitochondrial translation[J]. Nature Reviews. Molecular Cell Biology, 2011, 12(1): 14-20.
[16] ARTERO-CASTRO A, KONDOH H, FERNANDEZ-MARCOS P J, et al. Rplp1 bypasses replicative senescence and contributes to transformation[J]. Experimental Cell Research, 2009, 315(8): 1372-1383.
[17] CAMPOS R K, WIJERATNE H R S, SHAH P, et al. Ribosomal stalk proteins RPLP1 and RPLP2 promote biogenesis of flaviviral and cellular multi-pass transmembrane proteins[J]. Nucleic Acids Research, 2020, 48(17): 9872-9885.
[18] ISHII K, WASHIO T, UECHI T, et al.Characteristics and clustering of human ribosomal protein genes[J]. BMC Genomics, 2006, 7: 37.
[19] BEHRMANN E, LOERKE J, BUDKEVICH T V, et al.Structural snapshots of actively translating human ribosomes[J]. Cell, 2015, 161(4): 845-857.
[20] ABBAS W, KUMAR A, HERBEIN G.The eEF1A proteins: at the crossroads of oncogenesis, apoptosis, and viral infections[J]. Frontiers in Oncology, 2015, 5(75): 1-10
[21] KIM J, NAMKUNG W, YOON J S, et al.The role of translation elongation factor eEF1A in intracellular alkalinization-induced tumor cell growth[J]. Laboratory Investigation, 2009, 89(8): 867-874.
[22] SASIKUMAR A N, PEREZ W B, KINZY T G.The many roles of the eukaryotic elongation factor 1 complex[J]. Wiley Interdisciplinary Reviews. RNA, 2012, 3(4): 543-555.
[23] CAMBY I, LE MERCIER M, LEFRANC F, et al.Galectin-1: a small protein with major functions[J]. Glycobiology, 2006, 16(11): 137R-157R.
[24] RABINOVICH G A, TOSCANO M A.Turning ‘sweet’ on immunity: galectin-glycan interactions in immune tolerance and inflammation[J]. Nature Reviews. Immunology, 2009, 9(5): 338-352.
[25] BINART N, ORMANDY C J, KELLY P A.Mammary gland development and the prolactin receptor[J]. Advances in Experimental Medicine and Biology, 2000, 480: 85-92.
[26] CLEVENGER C V, KLINE J B.Prolactin receptor signal transduction[J]. Lupus, 2001, 10(10): 706-718.
[27] YAHATA T, SHAO W, ENDOH H, et al.Selective coactivation of estrogen-dependent transcription by CITED1 CBP/p300-binding protein[J]. Genes & Development, 2001, 15(19): 2598-2612.
[28] DELGOFFE G M, VIGNALI D A.A Fox of a different color: FoxA1 programs a new regulatory T cell subset[J]. Nature Medicine, 2014, 20(3): 236-237.
[29] SONG L, WEI X, ZHANG B, et al.Role of Foxa1 in regulation of bcl2 expression during oxidative-stress-induced apoptosis in A549 type Ⅱ pneumocytes[J]. Cell Stress & Chaperones, 2009, 14(4): 417-425.
[30] GUO Y H, GAO W, LI Q, et al.Tissue inhibitor of metalloproteinases-4 suppresses vascular smooth muscle cell migration and induces cell apoptosis[J]. Life Sciences, 2004, 75(20): 2483-2493.
[31] TUMMALAPALLI C M, HEATH B J, TYAGI S C.Tissue inhibitor of metalloproteinase-4 instigates apoptosis in transformed cardiac fibroblasts[J]. Journal of Cellular Biochemistry, 2001, 80(4): 512-521.
[32] NGUYEN T, LIU X K, ZHANG Y L, et al.BTNL2, a butyrophilin-like molecule that functions to inhibit T cell activation[J]. Journal of Immunology, 2006, 176(12): 7354-7360.
[33] SWANSON R M, GAVIN M A, ESCOBAR S S, et al.Butyrophilin-like 2 modulates B7 costimulation to induce Foxp3 expression and regulatory T cell development in mature T cells[J]. Journal of Immunology, 2013, 190(5): 2027-2035.
[34] GUENGERICH F P.Cytochrome P450 2E1 and its roles in disease[J]. Chemico-Biological Interactions, 2020, 322: 109056.
[35] HOXHAJ G, MANNING B D.The PI3K-AKT network at the interface of oncogenic signalling and cancer metabolism[J]. Nature Review·Cancer, 2020, 20(2): 74-88.
[36] MANNING B D, TOKER A.AKT/PKB Signaling: navigating the network[J]. Cell, 2017, 169(3): 381-405.
[37] LIU Y, ZHANG Y, JIANG J, et al.GHR/PRLR heteromultimer is composed of GHR homodimers and PRLR homodimers[J]. Molecular Endocrinology, 2016, 30(5): 504-517.
[38] XU J, SUN D, JIANG J, et al.The role of prolactin receptor in GH signaling in breast cancer cells[J]. Molecular Endocrinology, 2013, 27(2): 266-279.
[39] TURNER N, GROSE R.Fibroblast growth factor signalling: from development to cancer[J]. Nature Reviews·Cancer, 2010, 10(2): 116-129.
[40] LIU M X, JIN L, SUN S J, et al.Metabolic reprogramming by PCK1 promotes TCA cataplerosis, oxidative stress and apoptosis in liver cancer cells and suppresses hepatocellular carcinoma[J]. Oncogene, 2018, 37(12): 1637-1653.
[41] DRIER Y, COTTON M J, WILLIAMSON K E, et al.An oncogenic MYB feedback loop drives alternate cell fates in adenoid cystic carcinoma[J]. Nature Genetics, 2016, 48(3): 265-272.
[42] GAUTAM S, FIORAVANTI J, ZHU W, et al.The transcription factor c-Myb regulates CD8(+) T cell stemness and antitumor immunity[J]. Nature Immunology, 2019, 20(3): 337-349.
[43] CAI Z N, LI W, MEHMOOD S, et al.Structural characterization, in vitro and in vivo antioxidant activities of a heteropolysaccharide from the fruiting bodies of Morchella esculenta[J]. Carbohydrate Polymers, 2018, 195: 29-38.
[44] LI W, CAI Z N, MEHMOOD S, et al.Polysaccharide FMP-1 from Morchella esculenta attenuates cellular oxidative damage in human alveolar epithelial A549 cells through PI3K/AKT/Nrf2/HO-1 pathway[J]. International Journal of Biological Macromolecules, 2018, 120(Pt A): 865-875.
[45] LI W, CAI Z N, MEHMOOD S, et al.Anti-inflammatory effects of Morchella esculenta polysaccharide and its derivatives in fine particulate matter-treated NR8383 cells[J]. International Journal of Biological Macromolecules, 2019, 129: 904-915.
[46] LI S, GAO A, DONG S, et al.Purification, antitumor and immunomodulatory activity of polysaccharides from soybean residue fermented with Morchella esculenta[J]. International Journal of Biological Macromolecules, 2017, 96:26-34.
[47] 李敬, 周伟杰, 赵士豪, 等. 羊肚菌多糖的分离纯化及其体外抗氧化、抗肿瘤活性研究[J]. 天然产物研究与开发, 2016, 28: 1351-1356.
[48] 陈彦, 潘见, 周丽伟, 等. 羊肚菌胞外多糖抗肿瘤作用的研究[J]. 食品科学, 2008, 29(9): 553-556.
[49] 蔡伟, 罗益远, 刘姝, 等. 南方红豆杉总多糖增强紫杉醇对S180荷瘤小鼠肿瘤抑制作用机制研究[J]. 现代中药研究与实践, 2021, 35(3): 38-42.