CEEPC/IPM/CMSC - Abstrakt prezentace

(Česká konference hmotnostní spektrometrie 2017 - P-021)
Radiosensitization of non-small cell lung cancer cells by autophagy inhibitor Lys05: phosphoproteomic analysis

Martin Ondrej 1 *, Lucie Čecháková 1, Ivo Fabrik 2, Barbora Šalovská 1,3, Aleš Tichý 1

  1. Department of Radiobiology, Faculty of Military Health Sciences in Hradec Králové
  2. Department of Molecular Pathology and Biology, Faculty of Military Health Sciences in Hradec Králové
  3. Institute of Medical Biochemistry, Faculty of Medicine in Hradec Králové

Abstrakt

Autophagy is a basic cellular process, which enables cells to adapt and survive in stress conditions, including exposure to ionizing radiation. By Recycling of macromolecules cells are able to obtain energy and keep homeostasis. Hence, autophagy is believed to represent a radioprotective mechanism. This radioprotective mechanism is largely utilized by cancer cells during radiotherapy leading to radioresistance. Inhibition of the cytoprotective autophagy seems to be a reasonable approach to sensitize cancer cells. In this respect, several inhibitors of autophagy have been synthesized, but radiosensitizing mechanism has not been fully understood yet.

In order to characterize the radiosensitizing mechanism, we employed SILAC approach based on quantitative phosphoproteomics. Non-small cell lung cancer cells H1299 (representative cancer treated by radiotherapy) were selected as a model cell line in order to study the effect of Lys05, a newly synthesized inhibitor of autophagy. The experimental workflow was based on shotgun proteomics with subsequent enrichment for phosphopeptides using TiO2 microspheres followed by final identification and quantification of proteins/phosphosites using LC-MS/MS. In total, more than 1,100 proteins were identified with more than 6,000 phosphosites. Proteins that were significantly modified were mostly localized in nucleus with role in regulation of macromolecule metabolic and biosynthetic processes, as well as in cellular response to stress. In addition, regulation of mTOR pathway (the crucial signaling associated with activation of autophagy) was observed suggesting one of the possible mechanisms involved in lung cancer cells radiosensitized by Lys05. Future work will be focused on description of biological processes and further validation.

* Korespondující autor: martin.ondrej@unob.cz

Poděkování:

The work was supported by Ministry of Defence, Czech republic (project: Long-term organization development plan 1011).


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