MicroRNA-101-3p suppresses proliferation and migration in hepatocellular carcinoma by targeting the HGF/c-Met pathway

  • Yang Liu
  • Juan Tan
  • Shuangyan Ou
  • Jun Chen
  • Limin ChenEmail author


MicroRNAs are involved in each stage of tumor development. Activation of the hepatocyte growth factor (HGF)/c-Met axis facilitates the proliferation and migration of cancer cells, and the HGF/c-MET pathway provides potential targets for anticancer treatment. However, the interaction between HGF and miRNAs in hepatocellular carcinoma (HCC) remains unknown. Previous studies have shown that miR-101 is downregulated in various types of cancer and acts as a tumor suppressor, but the role of miR-101 in HCC has not yet been well defined. Here, we show that HGF is upregulated while microRNA-101-3p is significantly downregulated in the tumor tissues of HCC. By combining bioinformatics analysis and luciferase reporter assays, we demonstrated that HGF is a direct target of miR-101. In vitro experiments indicated that miR-101 inhibits the migration and proliferation of HCC cells by targeting the HGF/c-MET axis, and in vivo studies showed that overexpressed miR-101 dramatically suppresses tumor growth. Therefore, the present study identifies miR-101 as a negative regulator of HGF/c-MET and suggests that miRNAs can be used as targeted drugs for the clinical treatment of HCC.


Hepatocellular carcinoma miR-101 HGF c-MET 



This study was supporting by Hunan Natural Science Youth Fund Project (Project number:2018JJ3784. Project name:Molecular mechanism of miR-1181 involvement in promoting proliferation and survival of hepatocellular). The funder had no role in study design; collection, analysis, or interpretation of data; in the writing of the report; or in the decision to submit this article for publication.

Compliance with ethical standards

Conflict of interests

Yang Liu declares that she has no conflict of interest. Juan Tan declares that she has no conflict of interest. Shuangyan Ou declares that she has no conflict of interest. Jun Chen declares that he has no conflict of interest. Limin Chen declares that she has no conflict of interest.

Ethical approval

All applicable international, national, and institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.


  1. 1.
    Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM (2010) Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008. Int J Cancer 127:2893–2917CrossRefGoogle Scholar
  2. 2.
    Eggert T, Greten TF (2017) Current standard and future perspectives in non-surgical therapy for hepatocellular carcinoma. Digestion 96:1–4CrossRefGoogle Scholar
  3. 3.
    Aravalli RN, Steer CJ, Cressman EN (2008) Molecular mechanisms of hepatocellular carcinoma. Hepatology 48:2047–2063CrossRefGoogle Scholar
  4. 4.
    Matsumoto K, Nakamura T, Sakai K, Nakamura T (2008) Hepatocyte growth factor and met in tumor biology and therapeutic approach with NK4. Proteomics 8:3360–3370CrossRefGoogle Scholar
  5. 5.
    Tokunou M, Niki T, Eguchi K, Iba S, Tsuda H, Yamada T, Matsuno Y, Kondo H, Saitoh Y, Imamura H, Hirohashi S (2001) c-MET expression in myofibroblasts: role in autocrine activation and prognostic significance in lung adenocarcinoma. Am J Pathol 158:1451–1463CrossRefGoogle Scholar
  6. 6.
    Mi J, Hooker E, Balog S, Zeng H, Johnson DT, He Y, Yu EJ, Wu H, Le V, Lee DH, Aldahl J, Gonzalgo ML, Sun Z (2018) Activation of hepatocyte growth factor/MET signaling initiates oncogenic transformation and enhances tumoraggressiveness in the murine prostate. J Biol Chem 293:20123–20136. CrossRefGoogle Scholar
  7. 7.
    Pisters LL, Troncoso P, Zhau HE, Li W, von Eschenbach AC, Chung LW (1995) c-met proto-oncogene expression in benign and malignant human prostate tissues. J Urol 154:293–298CrossRefGoogle Scholar
  8. 8.
    De Silva DM, Roy A, Kato T, Cecchi F, Lee YH, Matsumoto K, Bottaro DP (2017) Targeting the hepatocyte growth factor/met pathway in cancer. Biochem Soc Trans 45:855–870CrossRefGoogle Scholar
  9. 9.
    Garajova I, Giovannetti E, Biasco G, Peters GJ (2015) c-Met as a Target for Personalized Therapy. Transl Oncogenomics 7:13–31Google Scholar
  10. 10.
    Hu CT, Wu JR, Cheng CC, Wu WS (2017) The therapeutic targeting of HGF/c-met signaling in hepatocellular carcinoma: alternative approaches. Cancers (Basel) 9Google Scholar
  11. 11.
    Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281–297CrossRefGoogle Scholar
  12. 12.
    Chen S, Sun KX, Liu BL, Zong ZH, Zhao Y (2016) MicroRNA-505 functions as a tumor suppressor in endometrial cancer by targeting TGF-alpha. Mol Cancer 15:11CrossRefGoogle Scholar
  13. 13.
    Mitamura T, Watari H, Wang L, Kanno H, Kitagawa M, Hassan MK, Kimura T, Tanino M, Nishihara H, Tanaka S, Sakuragi N (2014) microRNA 31 functions as an endometrial cancer oncogene by suppressing Hippo tumor suppressor pathway. Mol Cancer 13:97CrossRefGoogle Scholar
  14. 14.
    Mirnezami AH, Pickard K, Zhang L, Primrose JN, Packham G (2009) MicroRNAs: key players in carcinogenesis and novel therapeutic targets. Eur J Surg Oncol 35:339–347CrossRefGoogle Scholar
  15. 15.
    Zhang H, Deng T, Liu R, Bai M, Zhou L, Wang X, Li S, Yang H, Li J, Ning T, Huang D, Li H, Zhang L, Ying G, Ba Y (2017) Exosome-delivered EGFR regulates liver microenvironment to promote gastric cancer liver metastasis. Nat Commun 8:15016CrossRefGoogle Scholar
  16. 16.
    Slaby O, Svoboda M, Michalek J, Vyzula R (2009) MicroRNAs in colorectal cancer: translation of molecular biology into clinical application. Mol Cancer 8:102CrossRefGoogle Scholar
  17. 17.
    Wang HJ, Ruan HJ, He XJ, Ma YY, Jiang XT, Xia YJ, Ye ZY, Tao HQ (2010) MicroRNA-101 is down-regulated in gastric cancer and involved in cell migration and invasion. Eur J Cancer 46:2295–2303CrossRefGoogle Scholar
  18. 18.
    Luo C, Merz PR, Chen Y, Dickes E, Pscherer A, Schadendorf D, Eichmuller SB (2013) MiR-101 inhibits melanoma cell invasion and proliferation by targeting MITF and EZH2. Cancer Lett 341:240–247CrossRefGoogle Scholar
  19. 19.
    Chen C, Ridzon DA, Broomer AJ, Zhou Z, Lee DH, Nguyen JT, Barbisin M, Xu NL, Mahuvakar VR, Andersen MR, Lao KQ, Livak KJ, Guegler KJ (2005) Real-time quantification of microRNAs by stem-loop RT-PCR. Nucleic Acids Res 33:e179CrossRefGoogle Scholar
  20. 20.
    Feng J, Wang K, Liu X, Chen S, Chen J (2009) The quantification of tomato microRNAs response to viral infection by stem-loop real-time RT-PCR. Gene 437:14–21CrossRefGoogle Scholar
  21. 21.
    Gong BD, Xie Q, Wang L, Xiang XG, Lin LY, Zhao GD, Wang H, Yu H (2009) Real-time quantification of microRNAs in Huh7 cells by stem-loop reverse transcriptase polymerase chain reaction. Zhonghua Gan Zang Bing Za Zhi 17:603–606Google Scholar
  22. 22.
    Shi Y, Yang F, Wei S, Xu G (2017) Identification of key genes affecting results of hyperthermia in osteosarcoma based on integrative ChIP-Seq/TargetScan analysis. Med Sci Monit 23:2042–2048CrossRefGoogle Scholar
  23. 23.
    Clements BA, Incani V, Kucharski C, Lavasanifar A, Ritchie B, Uludag H (2007) A comparative evaluation of poly-L-lysine-palmitic acid and Lipofectamine 2000 for plasmid delivery to bone marrow stromal cells. Biomaterials 28:4693–4704CrossRefGoogle Scholar
  24. 24.
    Dalby B, Cates S, Harris A, Ohki EC, Tilkins ML, Price PJ, Ciccarone VC (2004) Advanced transfection with Lipofectamine 2000 reagent: primary neurons, siRNA, and high-throughput applications. Methods 33:95–103CrossRefGoogle Scholar
  25. 25.
    Yan X, Liang H, Deng T, Zhu K, Zhang S, Wang N, Jiang X, Wang X, Liu R, Zen K, Zhang CY, Ba Y, Chen X (2013) The identification of novel targets of miR-16 and characterization of their biological functions in cancer cells. Mol Cancer 12:92CrossRefGoogle Scholar
  26. 26.
    Nora D, Kurashige Y, Shudo K, Takahashi A, Abiko Y, Saitoh M (2015) Effect of epithelial cells derived from periodontal ligament on osteoblast-like cells in a Transwell membrane coculture system. Arch Oral Biol 60:1007–1012CrossRefGoogle Scholar
  27. 27.
    Moya-Horno I, Viteri S, Karachaliou N, Rosell R (2018) Combination of immunotherapy with targeted therapies in advanced non-small cell lung cancer (NSCLC). Ther Adv Med Oncol 10:1758834017745012CrossRefGoogle Scholar
  28. 28.
    Blumenthal GM, Karuri SW, Zhang H, Zhang L, Khozin S, Kazandjian D, Tang S, Sridhara R, Keegan P, Pazdur R (2015) Overall response rate, progression-free survival, and overall survival with targeted and standard therapies in advanced non-small-cell lung cancer: US Food and Drug Administration trial-level and patient-level analyses. J Clin Oncol 33:1008–1014CrossRefGoogle Scholar
  29. 29.
    Puccini A, Marin-Ramos NI, Bergamo F, Schirripa M, Lonardi S, Lenz HJ, Loupakis F, Battaglin F (2019) Safety and tolerability of c-MET inhibitors in Cancer. Drug Saf 42:211–233. CrossRefGoogle Scholar
  30. 30.
    Hughes VS, Siemann DW (2019) Failures in preclinical and clinical trials of c-Met inhibitors: evaluation of pathway activity as a promising selection criterion. Oncotarget 10:184–197CrossRefGoogle Scholar
  31. 31.
    Zhu Q, Gong L, Wang J, Tu Q, Yao L, Zhang JR, Han XJ, Zhu SJ, Wang SM, Li YH, Zhang W (2016) miR-10b exerts oncogenic activity in human hepatocellular carcinoma cells by targeting expression of CUB and sushi multiple domains 1 (CSMD1). BMC Cancer 16:806CrossRefGoogle Scholar
  32. 32.
    Feng X, Jiang J, Shi S, Xie H, Zhou L, Zheng S (2016) Knockdown of miR-25 increases the sensitivity of liver cancer stem cells to TRAIL-induced apoptosis via PTEN/PI3K/Akt/Bad signaling pathway. Int J Oncol 49:2600–2610CrossRefGoogle Scholar
  33. 33.
    Zhang JJ, Wang CY, Hua L, Yao KH, Chen JT, Hu JH (2015) miR-107 promotes hepatocellular carcinoma cell proliferation by targeting Axin2. Int J Clin Exp Pathol 8:5168–5174Google Scholar
  34. 34.
    Kabir TD, Ganda C, Brown RM, Beveridge DJ, Richardson KL, Chaturvedi V, Candy P, Epis M, Wintle L, Kalinowski F, Kopp C, Stuart LM, Yeoh GC, George J, Leedman PJ (2018) A microRNA-7/growth arrest specific 6/TYRO3 axis regulates the growth and invasiveness of sorafenib-resistant cells in human hepatocellular carcinoma. Hepatology 67:216–231CrossRefGoogle Scholar
  35. 35.
    He R, Yang L, Lin X, Chen X, Lin X, Wei F, Liang X, Luo Y, Wu Y, Gan T, Dang Y, Chen G (2015) MiR-30a-5p suppresses cell growth and enhances apoptosis of hepatocellular carcinoma cells via targeting AEG-1. Int J Clin Exp Pathol 8:15632–15641Google Scholar
  36. 36.
    Jin Y, Wang J, Han J, Luo D, Sun Z (2017) MiR-122 inhibits epithelial-mesenchymal transition in hepatocellular carcinoma by targeting Snail1 and Snail2 and suppressing WNT/beta-cadherin signaling pathway. Exp Cell Res 360:210–217CrossRefGoogle Scholar
  37. 37.
    Xu Y, An Y, Wang Y, Zhang C, Zhang H, Huang C, Jiang H, Wang X, Li X (2013) miR-101 inhibits autophagy and enhances cisplatin-induced apoptosis in hepatocellular carcinoma cells. Oncol Rep 29:2019–2024CrossRefGoogle Scholar
  38. 38.
    Xie Y, Yao Q, Butt AM, Guo J, Tian Z, Bao X, Li H, Meng Q, Lu J (2014) Expression profiling of serum microRNA-101 in HBV-associated chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. Cancer Biol Ther 15:1248–1255CrossRefGoogle Scholar
  39. 39.
    Liu LN, Li DD, Xu HX, Zheng SG, Zhang XP (2015) Role of microRNAs in hepatocellular carcinoma. Front Biosci (Landmark Ed) 20:1056–1067CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Pathology, Infectious Diseases InstituteThe Third Xiangya Hospital of Central South UniversityChangshaChina
  2. 2.Medical Oncology Institute, Hunan Cancer HospitalChangshaChina
  3. 3.Hunan Polytechnic of Environment and BiologyHengyangChina

Personalised recommendations