PDF(4863 KB)
促进肝癌致癌活性和细胞代谢的新型介质——miR-516a-3p
Tao Rui, Xueyou Zhang, Shi Feng, Haitao Huang, Shaowei Zhan, Haiyang Xie, Lin Zhou, Shusen Zheng, Qi Ling
工程（英文） ›› 2022, Vol. 16 ›› Issue (9) : 162-175.
PDF(4863 KB)
PDF(4863 KB)
促进肝癌致癌活性和细胞代谢的新型介质——miR-516a-3p
MiR-516a-3p is a Novel Mediator of Hepatocellular Carcinoma Oncogenic Activity and Cellular Metabolism
肝细胞肝癌（HCC）是目前最致命的恶性肿瘤之一。根据先前的研究，19 号染色体miRNA簇（C19MC）与肝癌患者的肿瘤高负荷和不良预后相关。目前的研究旨在探讨miR-516a-3p 在HCC 中的作用。miR-516a-3p 是一种由C19MC 4 个致癌前体miRNA（即mir-516a-1、mir-516a-2、mir-516b-1 和mir-516b-2）所共同剪接而成的相同的成熟体miRNA。在肝癌队列中，与瘤旁组织相比，miR-516a-3p 在肝癌组织中显著高表达。肿瘤miR-516a-3p 的高表达与肝癌高肿瘤负荷相关，可以区分高HCC复发率和死亡率，并独立预测肝癌的不良预后。进一步通过体外实验发现miR-516a-3p 增强了肝癌细胞的增殖、迁移和侵袭性，并通过体内实验验证miR-516a-3p 促进了肿瘤的增殖和远处转移能力。在肝癌细胞中，miR-516a-3p 可以通过外泌体进行递送，并增加受体肝癌细胞的致癌活性。此外，为探索miR-516a-3p 致癌的潜在机制，本研究进行了全面的转录组学、蛋白质组学和代谢组学分析。多组学DIABLO分析显示，蛋白质组学和代谢组学数据之间具有密切的相关性和较强的聚类一致性。进一步证实了6 种基因的mRNA（即LMBR1、CHST9、RBM3、SLC7A6、PTGFRN和NOL12）是miR-516a-3p 的直接靶点，并在miR-516a-3p 介导的代谢调节中发挥核心作用。综合多组学和共富集途径分析表明，miR-516a-3p 可以调节肝癌细胞的代谢途径，特别是嘌呤代谢和嘧啶代谢。总之，本研究发现，miR-516a-3p 可以通过调节细胞代谢和外泌体递送系统
影响相邻细胞，促进肝癌细胞的肿瘤恶性进展。因此，miR-516a-3p可作为肝癌治疗的新分子靶点。
Hepatocellular carcinoma (HCC) remains one of the most lethal malignancies. We previously demonstrated that the chromosome 19 microRNA cluster (C19MC) was associated with tumor burden and prognosis in patients with HCC. In the current study, we aim to explore the role of miR-516a-3p—an identical mature microRNA (miRNA) co-spliced by four oncogenic pre-miRNAs of C19MC (i.e., mir-516a-1, mir-516a-2, mir-516b-1, and mir-516b-2)—in HCC. In our cohort of HCC patients, miR-516a-3p was highly expressed in HCC tissues in comparison with adjacent non-tumor tissues. High expression of tumor miR-516a-3p significantly correlated with advanced tumor stages, distinguished high HCC recurrence and mortality, and independently predicted poor prognosis. We further found that miR-516a-3p enhanced the proliferation, migration, and invasiveness of HCC cells in vitro and promoted tumor growth and metastasis in vivo. Among cancer cells, miR-516a-3p could be delivered via exosomes or extracellular vesicles and increased the oncogenic activity of recipient cells. Moreover, we performed comprehensive transcriptomics, proteomics, and metabolomics analysis on the potential mechanism underlying miR-516a-3p-promoted oncogenicity. MixOmic DIABLO analysis showed a close correlation and strong cluster consistency between the proteomics and metabolomics datasets. We further confirmed six proteins (i.e., LMBR1, CHST9, RBM3, SLC7A6, PTGFRN, and NOL12) as the direct targets of miR-516a-3p and as central players in miR-516a-3p-mediated metabolism regulation. The integrated multi-omics and co-enriched pathway analysis showed that miR-516a-3p regulates the metabolic pathways of HCC cells, particularly purine and pyrimidine metabolism. In conclusion, our findings suggest that miR-516a-3p promotes malignant behaviors in HCC cells by regulating cellular metabolism and affecting neighboring cells via the exosome delivery system. Thus, we suggest miR-516a-3p as a novel molecular target for HCC therapy.
Hepatocellular carcinoma / miRNA cluster / Exosome / Multi-omics
| [1] |
Siegel RL, Miller KD, Jemal A. Cancer statistics, 2018. CA Cancer J Clin 2018; 68(1):7–30.
|
| [2] |
Cronin KA, Lake AJ, Scott S, Sherman RL, Noone AM, Howlader N, et al. Annual report to the nation on the status of cancer, part I: national cancer statistics. Cancer 2018;124(13):2785–800.
|
| [3] |
Ryder SD. Guidelines for the diagnosis and treatment of hepatocellular carcinoma (HCC) in adults. Gut 2003;52:iii1–8.
|
| [4] |
Bruix J, Qin S, Merle P, Granito A, Huang YH, Bodoky G, et al. Regorafenib for patients with hepatocellular carcinoma who progressed on sorafenib treatment (RESORCE): a randomised, double-blind, placebo-controlled, phase 3 trial. Lancet 2017;389(10064):56–66.
|
| [5] |
Livraghi T, Meloni F, Di Stasi M, Rolle E, Solbiati L, Tinelli C, et al. Sustained complete response and complications rates after radiofrequency ablation of very early hepatocellular carcinoma in cirrhosis: is resection still the treatment of choice? Hepatology 2008;47(1):82–9.
|
| [6] |
Bruix J, Sala M, Llovet JM. Chemoembolization for hepatocellular carcinoma. Gastroenterology 2004;127(5 Suppl 1):S179–88.
|
| [7] |
Sapisochin G, Bruix J. Liver transplantation for hepatocellular carcinoma: outcomes and novel surgical approaches. Nat Rev Gastroenterol Hepatol 2017;14(4):203–17.
|
| [8] |
Berindan-Neagoe I, Monroig PDC, Pasculli B, Calin GA. MicroRNAome genome: a treasure for cancer diagnosis and therapy. CA Cancer J Clin 2014;64(5): 311–36.
|
| [9] |
Wang Y, Luo J, Zhang H, Lu J. microRNAs in the same clusters evolve to coordinately regulate functionally related genes. Mol Biol Evol 2016;33(9): 2232–47.
|
| [10] |
Kabekkodu SP, Shukla V, Varghese VK, D’ Souza J, Chakrabarty S, Satyamoorthy K. Clustered miRNAs and their role in biological functions and diseases. Biol Rev 2018;93(4):1955–86.
|
| [11] |
Gennarino VA, D’Angelo G, Dharmalingam G, Fernandez S, Russolillo G, Sanges R, et al. Identification of microRNA-regulated gene networks by expression analysis of target genes. Genome Res 2012;22(6):1163–72.
|
| [12] |
Pagliuca A, Valvo C, Fabrizi E, di Martino S, Biffoni M, Runci D, et al. Analysis of the combined action of miR-143 and miR-145 on oncogenic pathways in colorectal cancer cells reveals a coordinate program of gene repression. Oncogene 2013;32(40):4806–13.
|
| [13] |
Noguer-Dance M, Abu-Amero S, Al-Khtib M, Lefevre A, Coullin P, Moore GE, et al. The primate-specific microRNA gene cluster (C19MC) is imprinted in the placenta. Hum Mol Genet 2010;19(18):3566–82.
|
| [14] |
Rui T, Xu S, Zhang X, Huang H, Feng S, Zhan S, et al. The chromosome 19 microRNA cluster, regulated by promoter hypomethylation, is associated with tumour burden and poor prognosis in patients with hepatocellular carcinoma. J Cell Physiol 2020;235(9):6103–12.
|
| [15] |
Augello C, Vaira V, Caruso L, Destro A, Maggioni M, Park YN, et al. MicroRNA profiling of hepatocarcinogenesis identifies C19MC cluster as a novel prognostic biomarker in hepatocellular carcinoma. Liver Int 2012;32(5): 772–82.
|
| [16] |
Fornari F, Milazzo M, Chieco P, Negrini M, Marasco E, Capranico G, et al. In hepatocellular carcinoma miR-519d is up-regulated by p53 and DNA hypomethylation and targets CDKN1A/p21, PTEN, AKT3 and TIMP2. J Pathol 2012;227(3):275–85.
|
| [17] |
Toffanin S, Hoshida Y, Lachenmayer A, Villanueva A, Cabellos L, Minguez B, et al. MicroRNA-based classification of hepatocellular carcinoma and oncogenic role of miR-517a. Gastroenterology 2011;140(5):1618–28.e16.
|
| [18] |
Mills J, Capece M, Cocucci E, Tessari A, Palmieri D. Cancer-derived extracellular vesicle-associated microRNAs in intercellular communication: one cell’s trash is another cell’s treasure. Int J Mol Sci 2019;20(24):6109.
|
| [19] |
Colombo M, Raposo G, Théry C. Biogenesis, secretion, and intercellular interactions of exosomes and other extracellular vesicles. Annu Rev Cell Dev Biol 2014;30(1):255–89.
|
| [20] |
Delorme-Axford E, Bayer A, Sadovsky Y, Coyne CB. Autophagy as a mechanism of antiviral defense at the maternal-fetal interface. Autophagy 2013;9(12): 2173–4.
|
| [21] |
Delorme-Axford E, Donker RB, Mouillet JF, Chu T, Bayer A, Ouyang Y, et al. Human placental trophoblasts confer viral resistance to recipient cells. Proc Natl Acad Sci USA 2013;110(29):12048–53.
|
| [22] |
Bullerdiek J, Flor I. Exosome-delivered microRNAs of ‘‘chromosome 19 microRNA cluster” as immunomodulators in pregnancy and tumorigenesis. Mol Cytogenet 2012;5(1):27.
|
| [23] |
Zhang X, Ye P, Huang H, Wang B, Dong F, Ling Q. TCF7L2 rs290487 C allele aberrantly enhances hepatic gluconeogenesis through allele-specific changes in transcription and chromatin binding. Aging 2020;12(13):13365–87.
|
| [24] |
Chong J, Soufan O, Li C, Caraus I, Li S, Bourque G, et al. MetaboAnalyst 4.0: towards more transparent and integrative metabolomics analysis. Nucleic Acids Res 2018;46(W1):W486–94.
|
| [25] |
Singh A, Shannon CP, Gautier B, Rohart F, Vacher M, Tebbutt SJ, et al. DIABLO: an integrative approach for identifying key molecular drivers from multiomics assays. Bioinformatics 2019;35(17):3055–62.
|
| [26] |
Karnovsky A, Weymouth T, Hull T, Tarcea VG, Scardoni G, Laudanna C, et al. Metscape 2 bioinformatics tool for the analysis and visualization of metabolomics and gene expression data. Bioinformatics 2012;28(3):373–80.
|
| [27] |
Rui T, Zhang X, Feng S, Huang H, Zhan S, Xie H, et al. The similar effects of miR512-3p and miR-519a-2-5p on the promotion of hepatocellular carcinoma: different tunes sung with equal skill. Front Oncol 2020;10:1244.
|
| [28] |
Meunier J, Lemoine F, Soumillon M, Liechti A, Weier M, Guschanski K, et al. Birth and expression evolution of mammalian microRNA genes. Genome Res 2013;23(1):34–45.
|
| [29] |
Heusermann W, Hean J, Trojer D, Steib E, von Bueren S, Graff-Meyer A, et al. Exosomes surf on filopodia to enter cells at endocytic hot spots, traffic within endosomes, and are targeted to the ER. J Cell Biol 2016;213(2):173–84.
|
| [30] |
Tian T, Wang Y, Wang H, Zhu Z, Xiao Z. Visualizing of the cellular uptake and intracellular trafficking of exosomes by live-cell microscopy. J Cell Biochem 2010;111(2):488–96.
|
| [31] |
Tseng LL, Lin CL, Tzen KY, Chang SC, Chang MF. LMBD1 protein serves as a specific adaptor for insulin receptor internalization. J Biol Chem 2013;288(45): 32424–32.
|
| [32] |
Shi H, Yao R, Lian S, Liu P, Liu Y, Yang YY, et al. Regulating glycolysis, the TLR4 signal pathway and expression of RBM3 in mouse liver in response to acute cold exposure. Stress 2019;22(3):366–76.
|
| [33] |
Orlicky DJ, Lieber JG, Morin CL, Evans RM. Synthesis and accumulation of a receptor regulatory protein associated with lipid droplet accumulation in 3T3-L1 cells. J Lipid Res 1998;39(6):1152–61.
|
| [34] |
Scott DD, Trahan C, Zindy PJ, Aguilar LC, Delubac MY, Van Nostrand EL, et al. Nol12 is a multifunctional RNA binding protein at the nexus of RNA and DNA metabolism. Nucleic Acids Res 2017;45(21):12509–28.
|
| [35] |
Liu P, Ge M, Hu J, Li X, Che L, Sun K, et al. A functional mammalian target of rapamycin complex 1 signaling is indispensable for c-Myc-driven hepatocarcinogenesis. Hepatology 2017;66(1):167–81.
|
| [36] |
Vander Heiden MG, DeBerardinis RJ. Understanding the intersections between metabolism and cancer biology. Cell 2017;168(4):657–69.
|
| [37] |
Villa E, Ali ES, Sahu U, Ben-Sahra I. Cancer cells tune the signaling pathways to empower de novo synthesis of nucleotides. Cancers 2019;11(5):688.
|
| [38] |
Yin J, Ren W, Huang X, Deng J, Li T, Yin Y. Potential mechanisms connecting purine metabolism and cancer therapy. Front Immunol 2018;9:1697.
|
| [39] |
Ma F, Zhu Y, Liu X, Zhou Q, Hong X, Qu C, et al. Dual-specificity tyrosine phosphorylation-regulated kinase 3 loss activates purine metabolism and promotes hepatocellular carcinoma progression. Hepatology 2019;70(5): 1785–803.
|
| [40] |
Santana-Codina N, Roeth AA, Zhang Yi, Yang A, Mashadova O, Asara JM, et al. Oncogenic KRAS supports pancreatic cancer through regulation of nucleotide synthesis. Nat Commun 2018;9(1):4945.
|
| [41] |
Feng X, Ma D, Zhao J, Song Y, Zhu Y, Zhou Q, et al. UHMK1 promotes gastric cancer progression through reprogramming nucleotide metabolism. Embo J 2020;39(5):e102541.
|
| [42] |
Newman AC, Maddocks ODK. One-carbon metabolism in cancer. Br J Cancer 2017;116(12):1499–504.
|
| [43] |
Ribbenstedt A, Ziarrusta H, Benskin JP, Schopfer FJ. Development, characterization and comparisons of targeted and non-targeted metabolomics methods. PLoS ONE 2018;13(11):e0207082.
|
| [44] |
Piazza I, Kochanowski K, Cappelletti V, Fuhrer T, Noor E, Sauer U, et al. A map of protein-metabolite interactions reveals principles of chemical communication. Cell 2018;172(1–2):358–72.e23.
|
/
| 〈 |
|
〉 |
