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Inhibition of Src tyrosine kinase reverts chemoresistance toward 5-fluorouracil in human pancreatic carcinoma cells: an involvement of epidermal growth factor receptor signaling

Oncogene volume 27, pages 7212–7222 (2008)Cite this article

Abstract

Resistance to chemotherapy is believed to be a major cause of treatment failure in pancreatic cancer. Thus, it is necessary to explore alternative therapeutic modalities to overcome drug resistance in pancreatic cancer treatment. We tested the hypothesis that Src tyrosine kinase inhibition could augment the chemosensitivity of 5-fluorouracil (5-FU)-resistant human pancreatic cancer cells to 5-FU. As detected by MTT proliferation assay, propidium iodide and annexin V staining, a combination of 5-FU+Src kinase inhibitor PP2 (4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine) reflected the chemotherapeutic sensitivity and restored the 5-FU-induced apoptosis in 5-FU-resistant cells. Furthermore, when small-interfering RNA approach to silence Src gene expression was applied, the degree of 5-FU-induced apoptosis was increased in all cell lines independently of the chemoresistance status. Western blotting and RT–PCR analysis revealed that the expression of thymidylate synthase (TS) was higher in 5-FU-resistant cells, however, decreased significantly after pretreatment with PP2. Furthermore, the combination of 5-FU+PP2 decreased the 5-FU-induced activation of epidermal growth factor receptor (EGFR)–AKT pathway. Finally, PP2 in combination with 5-FU substantially decreased the in vivo tumor growth and inhibited distant metastases. Taken together, 5-FU chemoresistance can be reversed through indirect TS regulation by inhibiting Src tyrosine kinase. A potential mechanism of action of Src kinase inhibitors on 5-FU chemosensitivity might be linked to the inhibition of 5-FU-induced EGFR–AKT activation.

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Acknowledgements

We thank AstraZeneca for providing with AZM475271. This research was supported by the grant of the clinical research group KFO 128/1-3 of the Deutsche Forschungsgemeinschaft (DFG) and the research grant GU 489/3-1 of the Deutsche Forschungsgemeinschaft (DFG) of the Schwerpunktprogramm ‘The tumor-vessel interface’. This report includes data that were generated during the doctoral thesis of Bettina Schwarz at the Medical School of the Ludwig-Maximilians-University, Munich.

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  1. I Ischenko and P Camaj: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Surgery, Munich University—Grosshadern Campus, Munich, Germany

    I Ischenko, P Camaj, H Seeliger, A Kleespies, M Guba, B Schwarz, C Graeb, M E Eichhorn, K-W Jauch & C J Bruns

  2. Department of Gastroenterology, Munich University—Grosshadern Campus, Munich, Germany

    E N De Toni

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  1. I Ischenko
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Correspondence to I Ischenko.

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Ischenko, I., Camaj, P., Seeliger, H. et al. Inhibition of Src tyrosine kinase reverts chemoresistance toward 5-fluorouracil in human pancreatic carcinoma cells: an involvement of epidermal growth factor receptor signaling. Oncogene 27, 7212–7222 (2008). https://doi.org/10.1038/onc.2008.326

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