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Pharmacologic inhibition of mTOR antagonizes the cytotoxic activity of pemetrexed in non-small cell lung cancer

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Abstract

Purpose

Pemetrexed, an inhibitor of thymidylate synthase (TS) and additional folate-dependent enzymes, is clinically active in patients suffering from “non-squamous” non-small cell lung cancer (NSCLC). High expression of TS has been implied as biomarker predictive of resistance to pemetrexed. Against this background, we studied whether inhibition of mTOR could lower expression of TS and thus sensitize NSCLC cells to pemetrexed.

Methods and results

Using squamous cell carcinoma and adenocarcinoma NSCLC cell lines, we observed that constitutive TS expression levels failed to correlate with sensitivity to growth inhibition or apoptosis imposed by pemetrexed in vitro. Interestingly, pemetrexed strongly induced TS RNA and protein expression in all cell lines. The allosteric “rapalogue” mTOR inhibitor everolimus suppressed constitutive, but not pemetrexed-induced TS expression. Surprisingly, cotreatment with everolimus protected NSCLC cells against pemetrexed-induced apoptosis. This resulted in increased long-term clonogenic survival of NSCLC cells treated with pemetrexed plus everolimus as compared to pemetrexed alone. No such negative interaction was observed when everolimus was combined with recombinant TRAIL, a proliferation-independent proapoptotic agent.

Conclusions

Rapalogues may suppress the antitumor activity of pemetrexed by slowing cell cycle progression. This should be considered when combining pemetrexed and mTOR inhibitors in NSCLC treatment.

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Acknowledgments

We thank all members of the Laboratory of Molecular Oncology, Department of Medical Oncology, West German Cancer Center, for their help and stimulating discussions. This work was supported by grants from the Deutsche Forschungsgemeinschaft (SCHU1541/5-1) and Deutsche Krebshilfe (No. 107993).

Conflict of interest

Stephan Herbertz is an employee of Novartis Pharma; Martin Schuler has served as consultant to and has received research support from Novartis.

Author information

Author notes

  1. Patricia S. Hähnel

    Present address: Department of Medicine III, Johannes Gutenberg University, Mainz, Germany

Authors and Affiliations

  1. Department of Medical Oncology, West German Cancer Center, University Hospital Essen, University Duisburg-Essen, 45122, Essen, Germany

    Boyka Markova, Patricia S. Hähnel, Stefan Kasper, Martin Schuler & Frank Breitenbuecher

  2. Novartis Pharma, 90429, Nürnberg, Germany

    Stephan Herbertz

  3. Ruhrlandklinik, Division of Thoracic Oncology, University Duisburg-Essen, 45239, Essen, Germany

    Martin Schuler

  4. Department of Medical Oncology, West German Cancer Center, University Hospital Essen, Hufelandstrasse 55, 45147, Essen, Germany

    Martin Schuler

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  1. Boyka Markova
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Corresponding author

Correspondence to Martin Schuler.

Electronic supplementary material

Below is the link to the electronic supplementary material.

432_2011_1123_MOESM1_ESM.pdf

Supplemental Figure 1. Thymidylate synthase expression in lung cancer cells following treatment with everolimus. NSCLC cells were cultured in the absence or presence (+) of everolimus (10 nM) for 24 h. Cell lysates were analyzed by immunoblotting for the expression of thymidylate synthase (TS), S6 ribosomal protein phosphorylation (pS6) and actin as a loading control (PDF 344 kb)

432_2011_1123_MOESM2_ESM.pdf

Supplemental Figure 2. Everolimus treatment alters the cell cycle distribution of lung cancer cells. Cell cycle analyses of FaDu, NCl-H23 and A549 cells following 48 h of treatment with everolimus (10 nM) (PDF 113 kb)

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Markova, B., Hähnel, P.S., Kasper, S. et al. Pharmacologic inhibition of mTOR antagonizes the cytotoxic activity of pemetrexed in non-small cell lung cancer. J Cancer Res Clin Oncol 138, 545–554 (2012). https://doi.org/10.1007/s00432-011-1123-9

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  • DOI: https://doi.org/10.1007/s00432-011-1123-9

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