A panel of serum exosomal microRNAs as predictive markers for chemoresistance in advanced colorectal cancer

  • Guoying Jin
  • Yuhang Liu
  • Jia Zhang
  • Zehua Bian
  • Surui Yao
  • Bojian Fei
  • Leyuan Zhou
  • Yuan YinEmail author
  • Zhaohui HuangEmail author
Original Article



Chemoresistance is a common problem for cancer treatment worldwide. Circulating exosomal microRNAs (miRNAs) have been considered as promising biomarkers of cancers. However, few studies have assessed the relationship between serum/plasma exosomal microRNAs and chemoresistance in colorectal cancer (CRC).


Based on previous microarray analysis, we selected 30 miRNAs which are aberrantly expressed during CRC progression and then detected their expression levels in three pairs of oxaliplatin/5-fluorouracil-resistant CRC cell lines and the corresponding secreted exosomes. Six candidate exosomal miRNAs were identified for further evaluating potential value in predicting chemotherapeutic effect in advanced CRC patients. Finally, the molecular mechanisms of these miRNAs in drug resistance were explored by bioinformatics preliminarily.


We observed that the expression of 14 miRNAs was significantly higher in three drug-resistant CRC cells comparing with their parental cells. Among these miRNAs, miR-21-5p, miR-1246, miR-1229-5p, miR-135b, miR-425 and miR-96-5p are also up-regulated in exosomes from culture media of resistant cells. Clinical sample analysis confirmed that the expression levels of miR-21-5p, miR-1246, miR-1229-5p and miR-96-5p in serum exosomes were significantly higher in chemoresistant patients in contrast with chemosensitive controls. ROC curve showed that the combination of the four miRNAs had an area of under the curve (AUC) of 0.804 (P < 0.05). In addition, GO analysis and KEGG pathway analysis revealed that these miRNAs were enriched in PI3K-Akt signaling pathway, FoxO signaling pathway and autophagy pathway.


Our study demonstrates that a panel of serum exosomal miRNAs containing miR-21-5p, miR-1246, miR-1229-5p and miR-96-5p could significantly distinguish the chemotherapy-resistant group from advanced colorectal cancer patients. Targeting these miRNAs may promote chemosensitivity to oxaliplatin and 5-fluorouracil, and might be promising strategy for CRC treatment.


Colorectal cancer Chemoresistance Exosome MicroRNAs 



This study was partially supported by grants from the National Natural Science Foundation of China (81672328 and 81772636), Fundamental Research Funds for the Central Universities (NOJUSRP51619B and JUSRP51710A), Medical Key Professionals Program of Jiangsu Province (AF052141), Medical Youth Professionals Program of Jiangsu Province (QNRC2016162), Medical Innovation Team Program of Wuxi (CXTP003), National First-class Discipline Program of Food Science and Technology (JUFSTR20180101), Project of the Wuxi Health and Family Planning Commission (Z201806).

Compliance with ethical standards

Conflict of interest

No potential conflicts of interest were disclosed.

Supplementary material

280_2019_3867_MOESM1_ESM.tif (24.7 mb)
Supplementary material 1 (TIFF 25310 kb)
280_2019_3867_MOESM2_ESM.docx (45 kb)
Supplementary material 2 (DOCX 45 kb)


  1. 1.
    Siegel RL, Miller KD, Fedewa SA, Ahnen DJ, Meester RGS, Barzi A, Jemal A (2017) Colorectal cancer statistics, 2017. Cancer J Clin 67(3):177–193. CrossRefGoogle Scholar
  2. 2.
    Chen W (2015) Cancer statistics: updated cancer burden in China. Chin J Cancer Res 27(1):1. Google Scholar
  3. 3.
    Panczyk M (2014) Pharmacogenetics research on chemotherapy resistance in colorectal cancer over the last 20 years. World J Gastroenterol 20(29):9775–9827. CrossRefGoogle Scholar
  4. 4.
    Becerra AZ, Probst CP, Tejani MA, Aquina CT, González MG, Hensley BJ, Noyes K, Monson JR, Fleming FJ (2016) Evaluating the prognostic role of elevated preoperative carcinoembryonic antigen levels in colon cancer patients: results from the national cancer database. Ann Surg Oncol 23(5):1554–1561. CrossRefGoogle Scholar
  5. 5.
    Raposo G, Stoorvogel W (2013) Extracellular vesicles: exosomes, microvesicles, and friends. J Cell Biol 200(4):373–383. CrossRefGoogle Scholar
  6. 6.
    Hamam R, Ali AM, Alsaleh KA, Kassem M, Alfayez M, Aldahmash A, Alajez NM (2016) microRNA expression profiling on individual breast cancer patients identifies novel panel of circulating microRNA for early detection. Sci Rep 6:25997. CrossRefGoogle Scholar
  7. 7.
    He L, Hannon GJ (2004) MicroRNAs: small RNAs with a big role in gene regulation. Nat Rev Genet 5(7):522–531. CrossRefGoogle Scholar
  8. 8.
    Hoshino I, Matsubara H (2013) MicroRNAs in cancer diagnosis and therapy: from bench to bedside. Surg Today 43(5):467–478. CrossRefGoogle Scholar
  9. 9.
    Yin Y, Zhang B, Wang W, Fei B, Quan C, Zhang J, Song M, Bian Z, Wang Q, Ni S, Hu Y, Mao Y, Zhou L, Wang Y, Yu J, Du X, Hua D, Huang Z (2014) miR-204-5p inhibits proliferation and invasion and enhances chemotherapeutic sensitivity of colorectal cancer cells by downregulating RAB22A. Clin Cancer Res 20(23):6187–6199. CrossRefGoogle Scholar
  10. 10.
    Chen X, Ba Y, Ma L et al (2008) Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. Cell Res 18(10):997–1006. CrossRefGoogle Scholar
  11. 11.
    Krek A, Grün D, Poy MN, Wolf R, Rosenberg L, Epstein EJ, MacMenamin P, da Piedade I, Gunsalus KC, Stoffel M, Rajewsky N (2005) Combinatorial microRNA target predictions. Nat Genet 37(5):495–500. CrossRefGoogle Scholar
  12. 12.
    The Gene Ontology Consortium (2019) The Gene Ontology Resource: 20 years and still GOing strong. Nucl Acids Res 47(D1):D330–D338. CrossRefGoogle Scholar
  13. 13.
    Gauthier SA, Pérez-González R, Sharma A, Huang FK, Alldred MJ, Pawlik M, Kaur G, Ginsberg SD, Neubert TA, Levy E (2017) Enhanced exosome secretion in Down syndrome brain - a protective mechanism to alleviate neuronal endosomal abnormalities. Acta Neuropathol Commun 5(1):65. CrossRefGoogle Scholar
  14. 14.
    Yin Y, Song M, Gu B, Qi X, Hu Y, Feng Y, Liu H, Zhou L, Bian Z, Zhang J, Zuo X, Huang Z (2016) Systematic analysis of key miRNAs and related signaling pathways in colorectal tumorigenesis. Gene 578(2):177–184. CrossRefGoogle Scholar
  15. 15.
    Huang Z, Huang S, Wang Q, Liang L, Ni S, Wang L, Sheng W, He X, Du X (2011) MicroRNA-95 promotes cell proliferation and targets sorting Nexin 1 in human colorectal carcinoma. Can Res 71(7):2582–2589. CrossRefGoogle Scholar
  16. 16.
    Vidal SJ, Rodriguez-Bravo V, Galsky M, Cordon-Cardo C, Domingo-Domenech J (2014) Targeting cancer stem cells to suppress acquired chemotherapy resistance. Oncogene 33(36):4451–4463. CrossRefGoogle Scholar
  17. 17.
    Hage C, Rausch V, Giese N, Giese T, Schönsiegel F, Labsch S, Nwaeburu C, Mattern J, Gladkich J, Herr I (2013) The novel c-Met inhibitor cabozantinib overcomes gemcitabine resistance and stem cell signaling in pancreatic cancer. Cell Death Dis 4:e627. CrossRefGoogle Scholar
  18. 18.
    Chu S, Liu G, Xia P, Chen G, Shi F, Yi T, Zhou H (2017) miR-93 and PTEN: key regulators of doxorubicin-resistance and EMT in breast cancer. Oncol Rep 38(4):2401–2407. CrossRefGoogle Scholar
  19. 19.
    Ogata-Kawata H, Izumiya M, Kurioka D, Honma Y, Yamada Y, Furuta K, Gunji T, Ohta H, Okamoto H, Sonoda H, Watanabe M, Nakagama H, Yokota J, Kohno T, Tsuchiya N (2014) Circulating exosomal microRNAs as biomarkers of colon cancer. PloS One 9(4):e92921. CrossRefGoogle Scholar
  20. 20.
    Uratani R, Toiyama Y, Kitajima T, Kawamura M, Hiro J, Kobayashi M, Tanaka K, Inoue Y, Mohri Y, Mori T, Kato T, Goel A, Kusunoki M (2016) Diagnostic potential of cell-free and exosomal MicroRNAs in the identification of patients with high-risk colorectal adenomas. PLoS One 11(10):e0160722. CrossRefGoogle Scholar
  21. 21.
    Qin Y, Li L, Wang F, Zhou X, Liu Y, Yin Y, Qi X (2018) Knockdown of Mir-135b sensitizes colorectal cancer cells to oxaliplatin-induced apoptosis through increase of FOXO1. Cell Physiol Biochem 48(4):1628–1637. CrossRefGoogle Scholar
  22. 22.
    Shao L, Chen Z, Soutto M, Zhu S, Lu H, Romero-Gallo J, Peek R, Zhang S, El-Rifai W (2019) Helicobacter pylori-induced miR-135b-5p promotes cisplatin resistance in gastric cancer. FASEB J 33(1):264–274. CrossRefGoogle Scholar
  23. 23.
    Liu B, Liu Y, Zhao L, Pan Y, Shan Y, Li Y, Jia L (2017) Upregulation of microRNA-135b and microRNA-182 promotes chemoresistance of colorectal cancer by targeting ST6GALNAC2 via PI3K/AKT pathway. Mol Carcinog 56(12):2669–2680. CrossRefGoogle Scholar
  24. 24.
    Su W, Mo Y, Wu F, Guo K, Li J, Luo Y, Ye H, Guo H, Li D, Yang Z (2016) miR-135b reverses chemoresistance of non-small cell lung cancer cells by downregulation of FZD1. Biomed Pharmacother 84:123–129. CrossRefGoogle Scholar
  25. 25.
    Zhang L, Cao D, Tang L, Sun C, Hu Y (2016) A panel of circulating miRNAs as diagnostic biomarkers for screening multiple myeloma: a systematic review and meta-analysis. Int J Lab Hematol 38(6):589–599. CrossRefGoogle Scholar
  26. 26.
    Zhu M, Huang Z, Zhu D, Zhou X, Shan X, Qi LW, Wu L, Cheng W, Zhu J, Zhang L, Zhang H, Chen Y, Zhu W, Wang T, Liu P (2017) A panel of microRNA signature in serum for colorectal cancer diagnosis. Oncotarget 8(10):17081–17091. CrossRefGoogle Scholar
  27. 27.
    Köberle V, Pleli T, Schmithals C, Augusto Alonso E, Haupenthal J, Bönig H, Peveling-Oberhag J, Biondi RM, Zeuzem S, Kronenberger B, Waidmann O, Piiper A (2013) Differential stability of cell-free circulating microRNAs: implications for their utilization as biomarkers. PLoS One 8(9):e75184. CrossRefGoogle Scholar
  28. 28.
    Tan Z, Zheng H, Liu X, Zhang W, Zhu J, Wu G, Cao L, Song J, Wu S, Song L, Li J (2016) MicroRNA-1229 overexpression promotes cell proliferation and tumorigenicity and activates Wnt/β-catenin signaling in breast cancer. Oncotarget 7(17):24076–24087. Google Scholar
  29. 29.
    Guo P, Yu Y, Tian Z, Lin Y, Qiu Y, Yao W, Zhang L (2018) Upregulation of miR-96 promotes radioresistance in glioblastoma cells via targeting PDCD4. Int J Oncol 53(4):1591–1600. Google Scholar
  30. 30.
    Hasegawa S, Eguchi H, Nagano H, Konno M, Tomimaru Y, Wada H, Hama N, Kawamoto K, Kobayashi S, Nishida N, Koseki J, Nishimura T, Gotoh N, Ohno S, Yabuta N, Nojima H, Mori M, Doki Y, Ishii H (2014) MicroRNA-1246 expression associated with CCNG2-mediated chemoresistance and stemness in pancreatic cancer. Br J Cancer 111(8):1572–1580. CrossRefGoogle Scholar
  31. 31.
    Li XJ, Ren ZJ, Tang JH, Yu Q (2017) Exosomal MicroRNA MiR-1246 promotes cell proliferation, invasion and drug resistance by targeting CCNG2 in breast cancer. Cell Physiol Biochem 44(5):1741–1748. CrossRefGoogle Scholar
  32. 32.
    Lin SS, Peng CY, Liao YW, Chou MY, Hsieh PL, Yu CC (2018) miR-1246 targets CCNG2 to enhance cancer stemness and chemoresistance in oral carcinomas. Cancers 10:8. CrossRefGoogle Scholar
  33. 33.
    Gaudelot K, Gibier JB, Pottier N, Hémon B, Van Seuningen I, Glowacki F, Leroy X, Cauffiez C, Gnemmi V, Aubert S, Perrais M (2017) Targeting miR-21 decreases expression of multi-drug resistant genes and promotes chemosensitivity of renal carcinoma. Tumour Biol 39(7):1010428317707372. CrossRefGoogle Scholar
  34. 34.
    Krause M, Dubrovska A, Linge A, Baumann M (2017) Cancer stem cells: Radioresistance, prediction of radiotherapy outcome and specific targets for combined treatments. Adv Drug Deliv Rev 109:63–73. CrossRefGoogle Scholar
  35. 35.
    Papa A, Pandolfi PP (2019) The PTENPI3K axis in cancer. Biomolecules 9:4. CrossRefGoogle Scholar
  36. 36.
    Zhu WJ, Chen X, Wang YW, Liu HT, Ma RR, Gao P (2017) MiR-1268b confers chemosensitivity in breast cancer by targeting ERBB2-mediated PI3K-AKT pathway. Oncotarget 8(52):89631–89642. Google Scholar
  37. 37.
    Eze N, Lee JW, Yang DH, Zhu F, Neumeister V, Sandoval-Schaefer T, Mehra R, Ridge JA, Forastiere A, Chung CH, Burtness B (2019) PTEN loss is associated with resistance to cetuximab in patients with head and neck squamous cell carcinoma. Oral Oncol 91:69–78. CrossRefGoogle Scholar
  38. 38.
    Liu G, Zhou J, Dong M (2019) Down-regulation of miR-543 expression increases the sensitivity of colorectal cancer cells to 5-Fluorouracil through the PTEN/PI3K/AKT pathway. Biosci Rep 39:3. Google Scholar
  39. 39.
    Huang X, Li Z, Zhang Q, Wang W, Li B, Wang L, Xu Z, Zeng A, Zhang X, Zhang X, He Z, Li Q, Sun G, Wang S, Li Q, Wang L, Zhang L, Xu H, Xu Z (2019) Circular RNA AKT3 upregulates PIK3R1 to enhance cisplatin resistance in gastric cancer via miR-198 suppression. Mol Cancer 18(1):71. CrossRefGoogle Scholar
  40. 40.
    Moon JH, Hong SW, Kim JE, Shin JS, Kim JS, Jung SA, Ha SH, Lee S, Kim J, Lee DH, Park YS, Kim DM, Park SS, Hong JK, Kim DY, Kim EH, Jung J, Kim MJ, Kim SM, Deming DA, Kim K, Kim TW, Jin DH (2019) Targeting β-catenin overcomes MEK inhibition resistance in colon cancer with KRAS and PIK3CA mutations. Br J Cancer 5:6. Google Scholar
  41. 41.
    Onoda N, Nakamura M, Aomatsu N, Noda S, Kashiwagi S, Kurata K, Uchino S, Hirakawa K (2015) Significant cytostatic effect of everolimus on a gefitinib-resistant anaplastic thyroid cancer cell line harboring PI3KCA gene mutation. Mol Clin Oncol 3(3):522–526. CrossRefGoogle Scholar
  42. 42.
    Wang JJ, Yu JP (2019) miR-181a down-regulates MAP2K1 to enhance adriamycin sensitivity in leukemia HL-60 cells. Eur Rev Med Pharmacol Sci 23(6):2497–2504. Google Scholar
  43. 43.
    Xu M, Zhou K, Wu Y, Wang L, Lu S (2019) Linc00161 regulated the drug resistance of ovarian cancer by sponging microRNA-128 and modulating MAPK1. Mol Carcinog 58(4):577–587. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Guoying Jin
    • 1
    • 2
  • Yuhang Liu
    • 1
    • 2
  • Jia Zhang
    • 1
    • 2
  • Zehua Bian
    • 1
    • 2
  • Surui Yao
    • 1
    • 2
  • Bojian Fei
    • 3
  • Leyuan Zhou
    • 4
  • Yuan Yin
    • 1
    • 2
    Email author
  • Zhaohui Huang
    • 1
    • 2
    Email author
  1. 1.Wuxi Cancer InstituteAffiliated Hospital of Jiangnan UniversityWuxiChina
  2. 2.Cancer Epigenetics Program, Wuxi School of MedicineJiangnan UniversityWuxiChina
  3. 3.Department of Surgical OncologyAffiliated Hospital of Jiangnan UniversityWuxiChina
  4. 4.Department of Radiation OncologyThe Affiliated Hospital of Jiangnan UniversityWuxiChina

Personalised recommendations