Tumor Biology

, Volume 37, Issue 7, pp 8765–8773 | Cite as

Inhibition of miR-15b decreases cell migration and metastasis in colorectal cancer

  • Jian Li
  • Yuxiang Chen
  • Xiong Guo
  • Ling Zhou
  • Zeming Jia
  • Yaping Tang
  • Ling Lin
  • Weidong Liu
  • Caiping Ren
Original Article


Colorectal cancer (CRC) has a high prevalence and mortality rate. Biomarkers for predicting the recurrence of CRC are not clinically available. This study investigated the role of circulating miR-15b in the prediction of CRC recurrence and the associated mechanism. miR-15b levels in plasma and tissues were measured by real-time PCR. Metastasis suppressor-1 (MTSS1) and Klotho protein expression were detected by Western blot and immunohistochemistry. Invasion and migration of CRC tumor cells were measured by transwell plates. Liver metastasis was established by intraspleen injection of HCT116 cells. Plasma miR-15b levels were significantly higher in CRC patients than in healthy controls, in CRC patients with metastasis than in CRC patients without metastasis, and in CRC patients with recurrence than in CRC patients without recurrence in the 5-year follow-up. miR-15b level in CRC tumors was significantly higher than that in peritumoral tissues. High plasma miR-15b level and negative MTSS1 and Klotho expression in tumor tissues significantly correlated with poor survival. Inhibition of miR-15b activity by adenovirus carrying antimiR-15b sequence significantly increased MTSS1 and Klotho protein expression and subsequently decreased colony formation ability, invasion, and migration of HCT116 cells in vitro and liver metastasis of HCT116 tumors in vivo. In conclusion, high abundance of circulating miR-15b correlated with tumor metastasis, recurrence, and poor patient prognosis through downregulation of MTSS1 and Klotho protein expression.


Colorectal cancer Metastasis miR-15b Prognosis MTSS1 Klotho 



This study was supported by the National 863 Hi-tech Project of China (2007AA021803, 2007AA021901, 2007AA021809, 2007AA021811), the National Natural Science Foundation of China (81272972), National Basic Research Program of China (2010CB833605), Hunan Provincial Science and Technology Department (2010FJ4030), Incubation Program for National Natural Science Funds for Distinguished Young Scholar of Central South University (2010QYZD006), and Open-End Fund for the Valuable and Precision Instruments of Central South University.

Compliance with ethical standards

This study was approved by the Ethics Committee for Human Research of Xiangya Hospital. The animal protocol was preapproved by Central South University, and all experiments were performed in accordance to the animal care guidelines of the Chinese Council.

Conflicts of interest



  1. 1.
    Jorgensen ML, Young JM, Solomon MJ. Optimal delivery of colorectal cancer follow-up care: improving patient outcomes. Patient Relat Outcome Meas. 2015;6:127–38.PubMedPubMedCentralGoogle Scholar
  2. 2.
    Siegel R, Ma J, Zhaohui Z, Jemal A. Cancer statistics, 2014. CA Cancer J Clin. 2014;64:9–29.CrossRefPubMedGoogle Scholar
  3. 3.
    Siegel R, DeSantis C, Jemal A. Colorectal cancer statistics, 2014. CA Cancer J Clin. 2014;64:104–17.CrossRefPubMedGoogle Scholar
  4. 4.
    Bujko K, Glimelius B, Valentini V, Michalski W, Spalek M. Postoperative chemotherapy in patients with rectal cancer receiving preoperative radio(chemo)therapy: a meta-analysis of randomized trials comparing surgery ± a fluoropyrimidine and surgery + a fluoropyrimidine ± oxaliplatin. Eur J Surg Oncol. 2015;41:713–23.CrossRefPubMedGoogle Scholar
  5. 5.
    Buie WD, Attard JA. Follow-up recommendations for colon cancer. Clin Colon Rectal Surg. 2005;18:232–43.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Meyerhardt JA, Mangu PB, Flynn PJ, Korde L, Loprinzi CL, Minsky BD, et al. Follow-up care, surveillance protocol, and secondary prevention measures for survivors of colorectal cancer: American Society of Clinical Oncology clinical practice guideline endorsement. J Clin Oncol. 2013;31:4465–70.CrossRefPubMedGoogle Scholar
  7. 7.
    Barrett LW, Sue FS, Wilton SD. Regulation of eukaryotic gene expression by the untranslated gene regions and other non-coding elements. Cell Mol Life Sci. 2012;69:3613–34.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Friedman RC, Farh KK, Burge CB, Bartel DP. Most mammalian mRNAs are conserved targets of microRNAs. Genome Res. 2009;19:92–105.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Jansson MD, Lund AH. MicroRNA and cancer. Mol Oncol. 2012;6:590–610.CrossRefPubMedGoogle Scholar
  10. 10.
    Bertoli G, Cava C, Castiglioni I. MicroRNAs: new biomarkers for diagnosis, prognosis, therapy prediction and therapeutic tools for breast cancer. Theranostics. 2015;5:1122–43.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Hollis M, Nair K, Vyas A, Chaturvedi LS, Gambhir S, Vyas D. MicroRNAs potential utility in colon cancer: early detection, prognosis, and chemosensitivity. World J Gastroenterol. 2015;21:8284–92.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Mostert B, Sieuwerts AM, Martens JW, Sleijfer S. Diagnostic applications of cell-free and circulating tumor cell-associated miRNAs in cancer patients. Expert Rev Mol Diagn. 2011;11:259–75.PubMedGoogle Scholar
  13. 13.
    Fesler A, Jiang J, Zhai H, Ju J. Circulating microRNA testing for the early diagnosis and follow-up of colorectal cancer patients. Mol Diagn Ther. 2014;18:303–8.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Satzger I, Mattern A, Kuettler U, Weinspach D, Voelker B, Kapp A, et al. MicroRNA-15b represents an independent prognostic parameter and is correlated with tumor cell proliferation and apoptosis in malignant melanoma. Int J Cancer. 2010;126:2553–62.PubMedGoogle Scholar
  15. 15.
    Kedmi M, Ben-Chetrit N, Körner C, Mancini M, Ben-Moshe NB, Lauriola M, et al. EGF induces microRNAs that target suppressors of cell migration: miR-15b targets MTSS1 in breast cancer. Sci Signal. 2015;8:ra29.CrossRefPubMedGoogle Scholar
  16. 16.
    Ota T, Doi K, Fujimoto T, Tanaka Y, Ogawa M, Matsuzaki H, et al. KRAS up-regulates the expression of miR-181a, miR-200c and miR-210 in a three-dimensional-specific manner in DLD-1 colorectal cancer cells. Anticancer Res. 2012;32:2271–5.PubMedGoogle Scholar
  17. 17.
    Zhang X, Kon T, Wang H, Li F, Huang Q, Rabbani ZN, et al. Enhancement of hypoxia-induced tumor cell death in vitro and radiation therapy in vivo by use of small interfering RNA targeted to hypoxia-inducible factor-1alpha. Cancer Res. 2004;64:8139–42.CrossRefPubMedGoogle Scholar
  18. 18.
    Shivapurkar N, Weiner LM, Marshall JL, Madhavan S, Deslattes Mays A, Juhl H, et al. Recurrence of early stage colon cancer predicted by expression pattern of circulating microRNAs. PLoS One. 2014;9:e84686.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Xie F, Ye L, Ta M, Zhang L, Jiang WG. MTSS1: a multifunctional protein and its role in cancer invasion and metastasis. Front Biosci (Schol Ed). 2011;3:621–31.CrossRefPubMedGoogle Scholar
  20. 20.
    Kayser G, Csanadi A, Kakanou S, Prasse A, Kassem A, Stickeler E, et al. Downregulation of MTSS1 expression is an independent prognosticator in squamous cell carcinoma of the lung. Br J Cancer. 2015;112:866–73.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Zhang J, Tong Y, Ren L, Li CD. Expression of metastasis suppressor 1 in cervical carcinoma and the clinical significance. Oncol Lett. 2014;8:2145–9.PubMedPubMedCentralGoogle Scholar
  22. 22.
    Zhang S, Qi Q. MTSS1 suppresses cell migration and invasion by targeting CTTN in glioblastoma. J Neurooncol. 2015;121:425–31.CrossRefPubMedGoogle Scholar
  23. 23.
    Zhou L, Li J, Shao QQ, Guo JC, Liang ZY, Zhou WX et al. Expression and significances of MTSS1 in pancreatic cancer. Pathol Oncol Res 2015 Jul 22. [Epub ahead of print]Google Scholar
  24. 24.
    Xie B, Chen J, Liu B, Zhan J. Klotho acts as a tumor suppressor in cancers. Pathol Oncol Res. 2013;19:611–7.CrossRefPubMedGoogle Scholar
  25. 25.
    Camilli TC, Xu M, O'Connell MP, Chien B, Frank BP, Subaran S, et al. Loss of Klotho during melanoma progression leads to increased filamin cleavage, increased Wnt5A expression, and enhanced melanoma cell motility. Pigment Cell Melanoma Res. 2011;24:175–86.CrossRefPubMedGoogle Scholar
  26. 26.
    Doi S, Zou Y, Togao O, Pastor JV, John GB, Wang L, et al. Klotho inhibits transforming growth factor-beta1 (TGF-beta1) signaling and suppresses renal fibrosis and cancer metastasis in mice. J Biol Chem. 2011;286:8655–65.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Zheng X, Chopp M, Lu Y, Buller B, Jiang F. MiR-15b and miR-152 reduce glioma cell invasion and angiogenesis via NRP-2 and MMP-3. Cancer Lett. 2013;329:146–54.CrossRefPubMedGoogle Scholar
  28. 28.
    Zhao Z, Zhang L, Yao Q, Tao Z. miR-15b regulates cisplatin resistance and metastasis by targeting PEBP4 in human lung adenocarcinoma cells. Cancer Gene Ther. 2015;22:108–14.CrossRefPubMedGoogle Scholar
  29. 29.
    Xia L, Zhang D, Du R, Pan Y, Zhao L, Sun S, et al. miR-15b and miR-16 modulate multidrug resistance by targeting BCL2 in human gastric cancer cells. Int J Cancer. 2008;123:372–9.CrossRefPubMedGoogle Scholar
  30. 30.
    Xia H, Qi Y, Ng SS, Chen X, Chen S, Fang M, et al. MicroRNA-15b regulates cell cycle progression by targeting cyclins in glioma cells. Biochem Biophys Res Commun. 2009;380:205–10.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Jian Li
    • 1
  • Yuxiang Chen
    • 1
  • Xiong Guo
    • 1
  • Ling Zhou
    • 1
  • Zeming Jia
    • 1
  • Yaping Tang
    • 1
  • Ling Lin
    • 1
  • Weidong Liu
    • 2
  • Caiping Ren
    • 2
  1. 1.Hepatobiliary and Enteric Surgery Research Center, Xiangya HospitalCentral South UniversityChangshaPeople’s Republic of China
  2. 2.Cancer Research Institute, Collaborative Innovation Center for Cancer Medicine, Key Laboratory for Carcinogenesis of Chinese Ministry of Health, School of Basic Medical ScienceCentral South UniversityChangshaPeople’s Republic of China

Personalised recommendations