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Gene expression profiling of breast cancer cells in response to gemcitabine: NF-κB pathway activation as a potential mechanism of resistance

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Abstract

Gemcitabine is a nucleoside analog with clinical relevance in the treatment of several solid tumors, including breast carcinoma. In spite of its cytotoxic effect, clinical efficacy is impaired by the development of resistance. We performed gene expression analysis to shed light into the molecular mechanism of action of this drug in two breast cancer cell lines. Activation of genes related with cell cycle, cell growth and apoptosis (BNIP3L, CCNG2, DDIT4, TGFB2, TP53BP1, TP53INP1, and VEGF) was the main finding in the p53-wild type cell line MCF7, while the p53-non-functional cell line MDA-MB-231 was characterized by the regulation of NF-κB target genes (BIRC3, CXCL1/GRO1, IRAK2, TNF, TNFAIP and TRAF1). Genes consistently induced (ATF3, CCNG2, CDKN1A, EGR1, INSIG1, and MAF) or repressed (CCND1 and VGF) in both cell lines, were also found after gemcitabine treatment. In addition, MDA-MB-231 cells showed a higher basal and induced NF-κB transcriptional activity after treatment with gemcitabine. In comparison with gemcitabine, gene expression after 5-fluorouracil treatment showed essentially different profiles in both cell lines. This, in spite of using equitoxic concentrations producing similar effects on cell cycle. NF-κB transcriptional activity in MDA-MB-231 cells was dependent on IκB-alpha phosphorylation, as shown by functional experiments using the specific inhibitor BAY11-7082. Moreover, immunohistochemical analysis of clinical samples of breast carcinoma further validated the induction of NF-κB expression and IκB down-regulation upon neoadjuvant gemcitabine treatment. Thus, gene expression patterns, in vitro functional studies and analysis of tissue samples are in agreement with a role for NF-κB pathway in gemcitabine response. Together with the reported role for NF-κB in the induction of resistance to chemotherapy, our data gives support to clinical strategies combining gemcitabine with NF-κB inhibitors in breast cancer.

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Acknowledgments

This work was supported by Ministerio de Educación y Ciencia, Grant Reference PTR1995-0753-OP and Ministerio de Ciencia y Tecnología SAF2001-0065 and SAF2004-08258-C02-01. GMB is junior investigator of the Ramón y Cajal Program 2004.

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Authors and Affiliations

  1. Breast and Gynaecological Cancer Group, Molecular Pathology Programme, Spanish National Cancer Centre (CNIO), Madrid, Spain

    Héctor Hernández-Vargas, Socorro María Rodríguez-Pinilla, Mercedes Julián-Tendero, José Palacios & Gema Moreno-Bueno

  2. Oncology Service, Complejo Hospitalario Ciudad de Jaén, Jaén, Spain

    Pedro Sánchez-Rovira

  3. Pathology Service, Complejo Hospitalario, Ciudad de Jaén, Jaén, Spain

    Cristóbal Cuevas

  4. Oncology Service, Hospital Universitario Miguel Servet, Zaragoza, Spain

    Antonio Antón

  5. Pathology Service, Hospital Universitario Miguel Servet, Zaragoza, Spain

    Maria Jesus Ríos

  6. Spanish National Breast Cancer Research Group (GEICAM), Madrid, Spain

    Pedro Sánchez-Rovira & Antonio Antón

  7. Group of Pathologists associated to GEICAM (pGEICAM), Madrid, Spain

    Cristóbal Cuevas & Maria Jesus Ríos

Authors
  1. Héctor Hernández-Vargas
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  2. Socorro María Rodríguez-Pinilla
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  9. Gema Moreno-Bueno
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Correspondence to José Palacios or Gema Moreno-Bueno.

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Hernández-Vargas, H., Rodríguez-Pinilla, S.M., Julián-Tendero, M. et al. Gene expression profiling of breast cancer cells in response to gemcitabine: NF-κB pathway activation as a potential mechanism of resistance. Breast Cancer Res Treat 102, 157–172 (2007). https://doi.org/10.1007/s10549-006-9322-9

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  • DOI: https://doi.org/10.1007/s10549-006-9322-9

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