DOI QR코드

DOI QR Code

Next-generation sequencing analysis of exosomal microRNAs: Fusobacterium nucleatum regulates the expression profiling of exosomal microRNAs in human colorectal cancer cells

  • Yu, Mi Ra (Department of Oral Pathology, and BK21 PLUS Project, School of Dentistry, Pusan National University) ;
  • Kim, Hye Jung (Periodontal Disease Signaling Network Research Center (MRC), School of Dentistry, Pusan National University) ;
  • Kang, Ji Wan (Interdisciplinary Program of Genomic Science, Pusan National University) ;
  • Kim, Yun Hak (Department of Anatomy, School of Medicine, Pusan National University) ;
  • Park, Hae Ryoun (Department of Oral Pathology, and BK21 PLUS Project, School of Dentistry, Pusan National University)
  • 투고 : 2020.08.21
  • 심사 : 2020.09.15
  • 발행 : 2020.09.30

초록

Colon cancer is one of the most common malignant tumors, but there are still a few validated biomarkers of colon cancer. Exosome-mediated microRNAs (miRNAs) have been recognized as potential biomarkers in cancers, and miRNAs can regulate a variety of genes. Recently, Fusobacterium nucleatum was discovered in the tissues of human colon cancer patients. Its role in colon cancer was highlighted. F. nucleatum may contribute to the progression of colon cancer through the mechanism of exosome-mediated miRNAs transfer. However, the exosomal miRNAs regulation mechanism by F. nucleatum in colon cancer is not well known. Thus, we performed next-generation sequencing to investigate the overall pattern of exosomal miRNAs expression in the colon cancer cell culture supernatant. We have confirmed the alterations of various exosomal miRNAs. In addition, to investigate the function of exosomal miRNAs, a Kyoto Encyclopedia of Genes and Genomes analysis was performed on the target genes of changed miRNAs. Potential target genes were associated with a variety of signaling pathways, and one of these pathways was related to colorectal cancer. These findings suggested that F. nucleatum can alter exosomal miRNAs released from colorectal cancer cells. Furthermore, exosomal miRNAs altered by F. nucleatum could be potential biomarkers for the diagnosis and therapy of colon cancer.

키워드

참고문헌

  1. Ma J, Yang Y, Fu Y, Guo F, Zhang X, Xiao S, Zhu W, Huang Z, Zhang J, Chen J. PIAS3-mediated feedback loops promote chronic colitis-associated malignant transformation. Theranostics 2018;8:3022-37. doi: 10.7150/thno.23046.
  2. Zackular JP, Baxter NT, Chen GY, Schloss PD. Manipulation of the gut microbiota reveals role in colon tumorigenesis. mSphere 2015;1:e00001-15. doi: 10.1128/mSphere.00001-15.
  3. Sun CH, Li BB, Wang B, Zhao J, Zhang XY, Li TT, Li WB, Tang D, Qiu MJ, Wang XC, Zhu CM, Qian ZR. The role of Fusobacterium nucleatum in colorectal cancer: from carcinogenesis to clinical management. Chronic Dis Transl Med 2019;5:178-87. doi: 10.1016/j.cdtm.2019.09.001.
  4. Yang Z, Ji G. Fusobacterium nucleatum-positive colorectal cancer. Oncol Lett 2019;18:975-82. doi: 10.3892/ol.2019.10433.
  5. Yan X, Liu L, Li H, Qin H, Sun Z. Clinical significance of Fusobacterium nucleatum , epithelial-mesenchymal transition, and cancer stem cell markers in stage III/IV colorectal cancer patients. Onco Targets Ther 2017;10:5031-46. doi: 10.2147/OTT.S145949.
  6. Bracci L, Lozupone F, Parolini I. The role of exosomes in colorectal cancer disease progression and response to therapy. Cytokine Growth Factor Rev 2020;51:84-91. doi: 10.1016/j.cytogfr.2019.12.004.
  7. Michael A, Bajracharya SD, Yuen PS, Zhou H, Star RA, Illei GG, Alevizos I. Exosomes from human saliva as a source of microRNA biomarkers. Oral Dis 2010;16:34-38. doi: 10.1111/j.1601-0825.2009.01604.x.
  8. Taylor DD, Gercel-Taylor C. MicroRNA signatures of tumorderived exosomes as diagnostic biomarkers of ovarian cancer. Gynecol Oncol 2008;110:13-21. doi: 10.1016/j.ygyno.2008.04.033.
  9. Lafitte M, Lecointre C, Roche S. Roles of exosomes in metastatic colorectal cancer. Am J Physiol Cell Physiol 2019;317:C869-80. doi: 10.1152/ajpcell.00218.2019.
  10. Xiao Y, Zhong J, Zhong B, Huang J, Jiang L, Jiang Y, Yuan J, Sun J, Dai L, Yang C, Li Z, Wang J, Zhong T. Exosomes as potential sources of biomarkers in colorectal cancer. Cancer Lett 2020;476:13-22. doi: 10.1016/j.canlet.2020.01.033.
  11. Hornick NI, Huan J, Doron B, Goloviznina NA, Lapidus J, Chang BH, Kurre P. Serum exosome microRNA as a minimally-invasive early biomarker of AML. Sci Rep 2015;5:11295. doi: 10.1038/srep11295.
  12. Guelfi G, Cochetti G, Stefanetti V, Zampini D, Diverio S, Boni A, Mearini E. Next generation sequencing of urine exfoliated cells: an approach of prostate cancer microRNAs research. Sci Rep 2018;8:7111. doi: 10.1038/s41598-018-24236-y.
  13. Chevillet JR, Kang Q, Ruf IK, Briggs HA, Vojtech LN, Hughes SM, Cheng HH, Arroyo JD, Meredith EK, Gallichotte EN, Pogosova-Agadjanyan EL, Morrissey C, Stirewalt DL, Hladik F, Yu EY, Higano CS, Tewari M. Quantitative and stoichiometric analysis of the microRNA content of exosomes. Proc Natl Acad Sci U S A 2014;111:14888-93. doi: 10.1073/pnas.1408301111.
  14. O'Brien SJ, Carter JV, Burton JF, Oxford BG, Schmidt MN, Hallion JC, Galandiuk S. The role of the miR-200 family in epithelial-mesenchymal transition in colorectal cancer: a systematic review. Int J Cancer 2018;142:2501-11. doi: 10.1002/ijc.31282.
  15. Jenjaroenpun P, Kremenska Y, Nair VM, Kremenskoy M, Joseph B, Kurochkin IV. Characterization of RNA in exosomes secreted by human breast cancer cell lines using next-generation sequencing. PeerJ 2013;1:e201. doi: 10.7717/peerj.201.
  16. Zheng L, Lin S, Lv C. MiR-26a-5p regulates cardiac fibroblasts collagen expression by targeting ULK1. Sci Rep 2018;8:2104. doi: 10.1038/s41598-018-20561-4.
  17. Liu C, Li G, Yang N, Su Z, Zhang S, Deng T, Ren S, Lu S, Tian Y, Liu Y, Qiu Y. miR-324-3p suppresses migration and invasion by targeting WNT2B in nasopharyngeal carcinoma. Cancer Cell Int 2017;17:2. doi: 10.1186/s12935-016-0372-8.
  18. He Y, Yu D, Zhu L, Zhong S, Zhao J, Tang J. miR-149 in human cancer: a systemic review. J Cancer 2018;9:375-88. doi: 10.7150/jca.21044.
  19. Sheedy P, Medarova Z. The fundamental role of miR-10b in metastatic cancer. Am J Cancer Res 2018;8:1674-88.
  20. Sandoval-Borquez A, Polakovicova I, Carrasco-Veliz N, Lobos-Gonzalez L, Riquelme I, Carrasco-Avino G, Bizama C, Norero E, Owen GI, Roa JC, Corvalan AH. MicroRNA-335-5p is a potential suppressor of metastasis and invasion in gastric cancer. Clin Epigenetics 2017;9:114. doi: 10.1186/s13148-017-0413-8.
  21. Zhang Z, Che X, Yang N, Bai Z, Wu Y, Zhao L, Pei H. miR-135b-5p Promotes migration, invasion and EMT of pancreatic cancer cells by targeting NR3C2. Biomed Pharmacother 2017;96:1341-8. doi: 10.1016/j.biopha.2017.11.074.
  22. Zhou C, Lu Y, Li X. miR-339-3p inhibits proliferation and metastasis of colorectal cancer. Oncol Lett 2015;10:2842-8. doi: 10.3892/ol.2015.3661.
  23. Huang ZM, Ge HF, Yang CC, Cai Y, Chen Z, Tian WZ, Tao JL. MicroRNA-26a-5p inhibits breast cancer cell growth by suppressing RNF6 expression. Kaohsiung J Med Sci 2019;35:467-73. doi: 10.1002/kjm2.12085.
  24. Zhang HQ, Sun Y, Li JQ, Huang LM, Tan SS, Yang FY, Li H. The expression of microRNA-324-3p as a tumor suppressor in nasopharyngeal carcinoma and its clinical significance. Onco Targets Ther 2017;10:4935-43. doi: 10.2147/OTT.S144223.
  25. Pucci C, Martinelli C, Ciofani G. Innovative approaches for cancer treatment: current perspectives and new challenges. Ecancermedicalscience 2019;13:961. doi: 10.3332/ecancer.2019.961.
  26. Yu X, Wang D, Wang X, Sun S, Zhang Y, Wang S, Miao R, Xu X, Qu X. CXCL12/CXCR4 promotes inflammation-driven colorectal cancer progression through activation of RhoA signaling by sponging miR-133a-3p. J Exp Clin Cancer Res 2019;38:32. doi: 10.1186/s13046-018-1014-x.
  27. Zhang S, Yang Y, Weng W, Guo B, Cai G, Ma Y, Cai S. Fusobacterium nucleatum promotes chemoresistance to 5-fluorouracil by upregulation of BIRC3 expression in colorectal cancer. J Exp Clin Cancer Res 2019;38:14. doi: 10.1186/s13046-018-0985-y.
  28. Chen B, Xia Z, Deng YN, Yang Y, Zhang P, Zhu H, Xu N, Liang S. Emerging microRNA biomarkers for colorectal cancer diagnosis and prognosis. Open Biol 2019;9:180212. https://doi.org/10.1098/rsob.180212
  29. Rossi S, Di Narzo AF, Mestdagh P, Jacobs B, Bosman FT, Gustavsson B, Majoie B, Roth A, Vandesompele J, Rigoutsos I, Delorenzi M, Tejpar S. microRNAs in colon cancer: a roadmap for discovery. FEBS Lett 2012;586:3000-7. doi: 10.1016/j.febslet.2012.07.048.
  30. Pan Q, Zhao J, Li M, Liu X, Xu Y, Li W, Wu S, Su Z. Exosomal miRNAs are potential diagnostic biomarkers between malignant and benign thyroid nodules based on next-generation sequencing. Carcinogenesis 2020;41:18-24. doi: 10.1093/carcin/bgz160.
  31. Rubie C, Kruse B, Frick VO, Kolsch K, Ghadjar P, Wagner M, Grasser F, Wagenpfeil S, Glanemann M. Chemokine receptor CCR6 expression is regulated by miR-518a-5p in colorectal cancer cells. J Transl Med 2014;12:48. doi: 10.1186/1479-5876-12-48.
  32. Ren L, Li Y, Zhao Q, Fan L, Tan B, Zang A, Yang H. miR-519 regulates the proliferation of breast cancer cells via targeting human antigen R. Oncol Lett 2020;19:1567-76. doi: 10.3892/ol.2019.11230.
  33. Guo H, Zeng X, Li H, Guo Y, Wang T, Guo H, Zhu G, Wang L, Zhou H, Liu K, Chen X, Wang H, Zhao X, Su H, Li Y. Plasma miR-1273g-3p acts as a potential biomarker for early Breast Ductal Cancer diagnosis. An Acad Bras Cienc 2020;92:e20181203. doi: 10.1590/0001-3765202020181203.
  34. Sun Y, Xiong X, Wang X. RELA promotes hypoxia-induced angiogenesis in human umbilical vascular endothelial cells via LINC01693/miR-302d/CXCL12 axis. J Cell Biochem 2019;120:12549-58. doi: 10.1002/jcb.28521.