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Carbonic anhydrase-IX inhibition enhances the efficacy of hexokinase II inhibitor for hepatocellular carcinoma in a murine model

  • Eun Ju Cho
  • Su Jong YuEmail author
  • Kyungmin Kim
  • Heki Cho
  • Young Youn Cho
  • Yun Bin Lee
  • Jeong-Hoon Lee
  • Yoon Jun Kim
  • Hyewon Youn
  • Jung-Hwan Yoon
Article

Abstract

Hypoxic conditions, which large or infiltrative hypovascular tumors may encounter, also produce acidic environments. Carbonic anhydrase-IX (CA-IX), an enzyme involved in lowering pH, is overexpressed in hepatocellular carcinoma (HCC). In the present study, whether inhibition of CA-IX enhances the efficacy of a hexokinase II inhibitor in an in vivo murine model was examined and its prognostic implication in HCC patients was investigated. CA-IX expression was evaluated using quantitative real-time PCR and western blot analysis using human HCC cell lines. 3-bromopyruvate (3-BP), a hexokinase II inhibitor, and acetazolamide, a carbonic anhydrase inhibitor, were used to target hexokinase II and CA-IX in vitro and in vivo, respectively. A human HCC cell line (Huh-7) was tested as a subcutaneous tumor model in BALB/c nu/nu mice. The prognostic role of CA-IX was evaluated in the TCGA database. Quantitative real-time PCR and western blot analysis revealed that CA-IX expression was activated in the presence of 3-BP. Further analysis showed that introducing an additional stress by treating the orally active CA-IX inhibitor (acetazolamide) can synergistically increase the efficacy of 3-BP in vivo, which was confirmed using a mouse model. We also found that HCC patients with high CA-IX expression show poor overall survival in TCGA database. These results indicate CA-IX is a promising therapeutic target for enhancing the efficacy of 3-BP and can be a prognostic factor for HCC.

Keywords

Hepatocellular carcinoma Acetazolamide 3-BP Apoptosis Hypoxia 

Notes

Acknowledgements

This study was funded by grants from the SNUH Research Fund (No. 04-2011-0660) and the Liver Research Foundation of Korea.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.

Supplementary material

10863_2019_9788_MOESM1_ESM.docx (20 kb)
ESM 1 (DOCX 19 kb)

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Copyright information

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

Authors and Affiliations

  • Eun Ju Cho
    • 1
  • Su Jong Yu
    • 1
    Email author
  • Kyungmin Kim
    • 2
  • Heki Cho
    • 3
  • Young Youn Cho
    • 4
  • Yun Bin Lee
    • 1
  • Jeong-Hoon Lee
    • 1
  • Yoon Jun Kim
    • 1
  • Hyewon Youn
    • 2
  • Jung-Hwan Yoon
    • 1
  1. 1.Department of Internal Medicine and Liver Research InstituteSeoul National University College of MedicineSeoulRepublic of Korea
  2. 2.Department of Biomedical sciences, and Nuclear Medicine, Cancer Research InstituteSeoul National University College of MedicineSeoulRepublic of Korea
  3. 3.Department of Internal MedicineDongguk University Ilsan HospitalGoyangRepublic of Korea
  4. 4.Department of Internal MedicineChung-Ang University HospitalSeoulRepublic of Korea

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