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Learn MoreIn this study we used unbiased approach in the lung cancer and colon cell lines (A549 and HTC 116 respectively) to identify universal early transcriptomic signatures of C-1305 cytotoxicity, and to highlight novel pathways responsible for its biological activity. The data obtained with real time analysis was used to select appropriate doses for subsequent RNAseq and biochemical analysis. Furthermore, the RNA samples prior RNA-seq analysis were pre-verified for transcriptomic activation of apoptosis related pathways via qPCR . Since our real time analysis of cell growth have shown that 24 h exposure to C-1305 (at IC50 concentrations) is sufficient to significantly alter A549 and HTC 116 cells growth and to activate apoptosis-related transcriptional signals, we determined genome wide transcriptomic alterations in A549 and HTC 116 upon C-1305 treatment. In brief, A549 and HTC 116 cells were exposed to 3 M or 10 M of C-1305, respectively for 24h day after plating. Furthermore, since our data have shown that 24h exposure of noncancer cells epithelial lung cells 16HBE14o- to 3 M C-1305 resulted in minimal cellular damage and loss of viability, we have included this treatment of 16HBE14o- as a negative control in our analysis. Finally, since genomic instabilities were reported for A549 cells to avoid related artifacts, DMSO vehicle treated cells (controls) were obtained after 24 h and after 8 h of cell culture. Total RNA was extracted from the cells after 24h exposure to C-1305 and 24 h and/or 8 h exposure to vehicle and subjected to RNA sequencing. SOURCE: Rafal,A,Bartoszewski Medical University of Gdansk
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