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The Effects of Different mTOR Inhibitors in EGFR Inhibitor Resistant Colon Carcinoma Cells

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Pathology & Oncology Research

Abstract

Several monoclonal antibodies and inhibitors targeting signalling pathways are being used in personalised medicine. Anti-EGFR antibodies seem to be effective, however, therapy resistance often occurs in colon carcinoma cases. mTOR inhibitors (mTORIs) could have a potential role in the breakthrough of therapy resistance. The mTOR activity related protein expression patterns and the in vitro effects of EGFR inhibitors (EGFRIs), mTORIs and their combinations were studied in different colon carcinoma cell lines (with different genetic backgrounds). Alamar Blue test and flow cytometry were used to analyse the in vitro proliferation and apoptotic effects of cetuximab, gefitinib, cisplatin, rapamycin, PP242 and NVP-BEZ235. The expressions of mTOR activity related proteins (p-70S6K, p-S6, Rictor, p-mTOR, Raptor) were studied by Western blot, immunocytochemistry and Duolink staining. The EGFRI resistance of the studied colon carcinoma cell lines related to their known mutations were confirmed, neither gefitinib nor cetuximab inhibited the proliferation or induced apoptosis in vitro. Individual differences in Rictor and Raptor expressions were detected by Western blot and immunocytochemistry beside elevated mTOR activity of these different colon carcinoma cell lines. These expression patterns correlated to the mTORIs sensitivity differences, moreover, mTORIs could enhance the effects of EGFRIs and other in vitro treatments. Our results suggest that mTORI combinations could be helpful in both EGFRI and platinum-based therapy of colon carcinomas. Moreover, we suggest determining both mTOR complex activity and mutations in Akt/mTOR signalling pathways for selecting the appropriate mTORIs and patients in potential future combination treatments.

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Acknowledgements

The project was supported by OTKA project (K84262) and Semmelweis University Scientific and Innovation Founds (STIA-KF-17); and by Bolyai fellowship (590/2015 - Hungarian Academy of Sciences) of the corresponding author (A. Sebestyén). The related research works of Z.Hujber and T. Dankó were also supported by New National Excellence Programs (ÚNKP-17-3; ÚNKP-17-2).

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Author notes

  1. Tamás Sticz and Anna Molnár contributed equally to this work.

Authors and Affiliations

  1. 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, Üllői út 26, Budapest, H-1085, Hungary

    Tamás Sticz, Anna Molnár, Titanilla Dankó, Zoltán Hujber, Gábor Petővári, Noémi Nagy, László Kopper & Anna Sebestyén

  2. Department of Transplantation and Surgery, Semmelweis University, Üllői út 26, Budapest, H-1085, Hungary

    Gyula Végső

  3. Tumour Progression Research Group of Joint Research Organization of Hungarian Academy of Sciences and Semmelweis University, Üllői út 26, Budapest, H-1085, Hungary

    Anna Sebestyén

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  1. Tamás Sticz
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Correspondence to Anna Sebestyén.

Additional information

Highlights:

- Activity of mTOR complexes in colon carcinoma cells correlates to mTORI sensitivity

- mTORIs promote anti-tumour effects of EGFRIs in therapy resistant colon carcinomas

- Additional markers and agents related to mTOR activity for personalised treatments

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Sticz, T., Molnár, A., Dankó, T. et al. The Effects of Different mTOR Inhibitors in EGFR Inhibitor Resistant Colon Carcinoma Cells. Pathol. Oncol. Res. 25, 1379–1386 (2019). https://doi.org/10.1007/s12253-018-0434-4

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  • DOI: https://doi.org/10.1007/s12253-018-0434-4

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