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Prognostic relevance of carbonic anhydrase IX expression is distinct in various subtypes of breast cancer and its silencing suppresses self-renewal capacity of breast cancer cells

Cancer Chemotherapy and Pharmacology volume 75, pages 235–246 (2015)Cite this article

Abstract

Purpose

Carbonic anhydrase IX (CAIX) is a hypoxia-inducible enzyme with extracellular catalytic domain that is overexpressed in a variety of cancers including breast cancer and plays a crucial role in maintaining favourable intracellular pH and reducing extracellular pH. The purpose of the current study was to elucidate the prognostic significance of CAIX in the intrinsic subtypes of breast cancer and to characterise CAIX as a drug target in breast cancer.

Methods

The prognostic significance of CAIX mRNA expression was interrogated in a cohort of 3,455 breast tumours by using an online tool, Kaplan–Meier plotter. The functional effects of stable CAIX depletion by shRNA in three breast cancer cell lines—MDA-MB-231, MCF7 and SKBR-3, representing basal-like, luminal A and HER2+ subtypes, respectively—were studied by proliferation, invasion, clonal spheroid formation and chemosensitivity assays under normoxia and hypoxia. Finally, the effect of pharmacological CA inhibition alone or in the combination with doxorubicin on self-renewal was assessed by spheroid-forming assay.

Results

High CAIX mRNA expression was significantly associated with poor survival in patients with basal-like, luminal B and triple-negative breast cancer, but not luminal A and HER+ subtypes. Silencing of CAIX expression had no significant effect on the cell proliferation or viability upon treatment with doxorubicin in any of the cell lines studied, while it inhibited spheroid formation in hypoxic conditions. Furthermore, pharmacological inhibition of CAs using acetazolamide had a synergistic effect with doxorubicin on decreasing the spheroid-forming efficiency in MDA-MB-231 cells.

Conclusions

Inhibition of CAIX reduces the self-renewal capacity of breast cancer cells, and the combination of doxorubicin and CAIX inhibition is an attractive therapeutic strategy in basal-like and triple-negative breast cancer, which warrants further investigations.

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Acknowledgments

This study was supported by the Latvian National Research Programme BIOMEDICINE.

Conflict of interest

None.

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Affiliations

  1. Latvian Biomedical Research and Study Centre, Ratsupites Str 1, Riga, 1067, Latvia

    Lāsma Ivanova, Elīna Zandberga, Karīna Siliņa, Zane Kalniņa, Artūrs Ābols, Edgars Endzeliņš & Aija Linē

  2. Latvian Institute of Organic Synthesis, Riga, Latvia

    Ineta Vendina, Nadezhda Romanchikova & Pēteris Trapencieris

  3. Latvian Oncology Center, Riga Eastern Clinical University Hospital, Riga, Latvia

    Alinta Hegmane & Jānis Eglītis

  4. Faculty of Medicine, University of Latvia, Riga, Latvia

    Alinta Hegmane & Jānis Eglītis

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  1. Lāsma Ivanova
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  2. Elīna Zandberga
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Correspondence to Aija Linē.

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Lāsma Ivanova and Elīna Zandberga have contributed equally to this work.

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Ivanova, L., Zandberga, E., Siliņa, K. et al. Prognostic relevance of carbonic anhydrase IX expression is distinct in various subtypes of breast cancer and its silencing suppresses self-renewal capacity of breast cancer cells. Cancer Chemother Pharmacol 75, 235–246 (2015). https://doi.org/10.1007/s00280-014-2635-1

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Keywords

  • Carbonic anhydrase IX
  • Hypoxia
  • Breast cancer
  • Spheroid-forming assay
  • Self-renewal