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The MUC4 mucin mediates gemcitabine resistance of human pancreatic cancer cells via the Concentrative Nucleoside Transporter family

Oncogene volume 32, pages 1714–1723 (2013)Cite this article

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Abstract

The fluorinated analog of deoxycytidine, Gemcitabine (Gemzar), is the main chemotherapeutic drug in pancreatic cancer, but survival remains weak mainly because of the high resistance of tumors to the drug. Recent works have shown that the mucin MUC4 may confer an advantage to pancreatic tumor cells by modifying their susceptibility to drugs. However, the cellular mechanism(s) responsible for this MUC4-mediated resistance is unknown. The aim of this work was to identify the cellular mechanisms responsible for gemcitabine resistance linked to MUC4 expression. CAPAN-2 and CAPAN-1 adenocarcinomatous pancreatic cancer (PC) cell lines were used to establish stable MUC4-deficient clones (MUC4-KD) by shRNA interference. Measurement of the IC50 index using tetrazolium salt test indicated that MUC4-deficient cells were more sensitive to gemcitabine. This was correlated with increased Bax/BclXL ratio and apoptotic cell number. Expression of Equilibrative/Concentrative Nucleoside Transporter (hENT1, hCNT1/3), deoxycytidine kinase (dCK), ribonucleotide reductase (RRM1/2) and Multidrug-Resistance Protein (MRP3/4/5) was evaluated by quantitative RT–PCR (qRT–PCR) and western blotting. Alteration of MRP3, MRP4, hCNT1 and hCNT3 expression was observed in MUC4-KD cells, but only hCNT1 alteration was correlated to MUC4 expression and sensitivity to gemcitabine. Decreased activation of MAPK, JNK and NF-κB pathways was observed in MUC4-deficient cells, in which the NF-κB pathway was found to have an important role in both sensitivity to gemcitabine and hCNT1 regulation. Finally, and in accordance with our in vitro data, we found that MUC4 expression was conversely correlated to that of hCNT1 in tissues from patients with pancreatic adenocarcinoma. This work describes a new mechanism of PC cell resistance to gemcitabine, in which the MUC4 mucin negatively regulates the hCNT1 transporter expression via the NF-κB pathway. Altogether, these data point out to MUC4 and hCNT1 as potential targets to ameliorate the response of pancreatic tumors to gemcitabine treatment.

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Acknowledgements

We thank MH Gevaert and R Siminsky (Department of Histology, Faculty of Medicine, University of Lille 2) and the technical platform IFR114/IMPRT for flow cytometry (Dr N Jouy) and luciferase (AS Drucbert) analyses. Gemcitabine and oxaliplatin are a kind gift from Dr A Lansiaux (Centre Oscar Lambret, Université Lille Nord de France, Inserm UMR837, team 4, Lille, France). Cytarabine/aracytin ARA-C is a kind gift from Pr B Quesnel (Oncohaematology department, Centre Hospitalier Régional et Universitaire de Lille, Inserm UMR837, team 3, Lille, France). We thank Dr JL Desseyn (Inserm U995) for his help in designing MUC4 shRNA. Nicolas Skrypek is a recipient of a PhD fellowship of Centre Hospitalier Régional et Universitaire (CHRU) de Lille and Région Nord-Pas de Calais. Dr Nicolas Jonckheere is a recipient of postdoctoral fellowship from the Institut National du Cancer (INCa) and Ligue Nationale contre le Cancer (LNCC). This work is supported by a grant from la Ligue Nationale contre le Cancer (Equipe Labellisée Ligue 2010, IVS). Isabelle Van Seuningen is the recipient of a ‘Contrat Hospitalier de Recherche Translationnelle’/CHRT 2010, AVIESAN. The funders had no role in study design, data collection and analysis, decision to publish or in preparation of the paper.

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Authors and Affiliations

  1. Inserm, UMR837, Jean-Pierre Aubert Research Center, Lille Cedex, France

    N Skrypek, B Duchêne, M Hebbar, E Leteurtre, I van Seuningen & N Jonckheere

  2. Université Lille Nord de France, Lille Cedex, France

    N Skrypek, B Duchêne, M Hebbar, E Leteurtre, I van Seuningen & N Jonckheere

  3. Centre Hospitalier Régional et Universitaire de Lille, Lille Cedex, France

    N Skrypek, B Duchêne, I van Seuningen & N Jonckheere

  4. Department of Medical Oncology, Centre Hospitalier Régional et Universitaire de Lille, Lille, France

    M Hebbar

  5. Department of Pathology, Centre Hospitalier Régional et Universitaire de Lille, Centre de Biologie-Pathologie, Lille, France

    E Leteurtre

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  1. N Skrypek
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Correspondence to N Jonckheere.

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Skrypek, N., Duchêne, B., Hebbar, M. et al. The MUC4 mucin mediates gemcitabine resistance of human pancreatic cancer cells via the Concentrative Nucleoside Transporter family. Oncogene 32, 1714–1723 (2013). https://doi.org/10.1038/onc.2012.179

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