Autophagy inhibition through small-molecule inhibitors is one of the approaches to increasing the efficiency of radiotherapy of oncological patients. A new inhibitor, Lys05, with potential to accumulate within lysosomes and to block autophagy has been discovered recently. Several studies have described its chemosensitizing effects, but nothing is known about its impact in the context of ionizing radiation (IR). To investigate the mechanisms underlying its role in radiosensitization we employed radioresistant human non-small cell lung carcinoma cells (H1299, p53-negative) in combination with multiple approaches (cell biology techniques to reveal the phenotypes, and quantitative phosphoproteomics to comprehensively describe Lys05-induced changes in irradiated cells). In the presented study, we report for the first time that Lys05 could be utilized in combination with IR as a promising future strategy in eradication of lung cancer cells.
Overall design
H1299 cells were either irradiated or left unirradiated and both of these groups were either treated with Lys05 or spautin or left untreated. This results in six experimental groups per one time interval. The cells were sampled at time one, twenty-four and fourty-eight hours post treatment. In each time & treatment group were n=3 samples. There were 6 experimental groups * 3 times * 3 biological replicates = 54 samples. These samples were hybridized in two-color fashion (portion of the three biological replicates were labeled with Cy3, part with Cy5) but were analyzed as single channel (quantile normalization across all channels).
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