Project description:Control (DMSO 0.1%; v/v) and 10 M DRB18 were used to treated 5 million A549 lung cancer cells in vitro for 48 hours. The untargeted metabolomics analysis was performed on the cell lysates. The main objective of the study was to determine changes in metabolite abundances in lung cancer after treatment with DRB18, an inhibitor of glucose transporter proteins.
Project description:The statsitcal model, latent pathway identification analysis (LPIA), was implemented for the analysis of A549 lung carcinoma cells treated with geldanamycin. Control and treated samples were assayed with Affymetrix HG_U133_plus_2 arrays and analyzed using LPIA. LPIA looks for statistically signï¬cant evidence of dysregulation in a network of pathways constructed in a manner that explicitly links pathways through their common function in the cell. Geldanamycin (geld) is known to inhibit the molecular chaperone protein, Hsp90, and plays a role in preventing the malignant transformation and proliferation of healthy cells during oncogenesis. LPIA successfully identified pathways specific to geldanamycin effects at the gene transcription level. A549 lung carcinoma cells were allowed to adhere for 24h and further incubated in the presence of geldanamycin at a concentration equivalent to the determined IC50 of 40 nM or IC20 of 10 nM, or with vehicle DMSO (final concentration 0.4%). After 24h and 48 h, cells were harvested and total RNA was purified and processed for Affymetrix HG_U133_Plus_2.0 microarray analysis. Raw probe intensities were RMA-normalized and avereaged for three replicates for each condition. Probe sets for 54,120-annotated open reading frames were included in LPIA analysis.
Project description:Control (DMSO 0.1%; v/v) and 10 µM DRB18 were used to treated 5 million A549 lung cancer cells in vitro for 48 hours. The untargeted metabolomics analysis was performed on the cell lysates. The main objective of the study was to determine changes in metabolite abundances in lung cancer after treatment with DRB18, an inhibitor of glucose transporter proteins.
Project description:In 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 µM or 10 µM of C-1305, respectively for 24 h day after plating. Furthermore, since our data have shown that 24 h exposure of noncancer cells epithelial lung cells 16HBE14o- to 3 µM 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 24 h exposure to C-1305 and 24 h and/or 8 h exposure to vehicle and subjected to RNA sequencing.
Project description:To identify genes regulated by SPDEF or FOXA3 in A549 lung carcinoma cells, we analyzed the whole-transcriptomic mRNA profiles of A549 cells expressing SPDEF or FOXA3 based on RNA-seq.
Project description:Title: Prediction of cell cycle behaviour in response to DNA damage agents via transcriptomic approach <br/> Description: Human lung adenocarcinoma A549 cells were treated with vehicle (DMSO) or Etoposide (ETOP, 10 M-BM-5M ), Benzo(a)pyrene (BaP, 25 M-BM-5M ), Cryptolepine ( CLP, 2.5 M-BM-5M) and 2-Amino-1-methyl-6-phenylimidazo [4,5-b]pyridine (PhIP, 50 M-BM-5M) for 6 h, 16 h, 24 h, 48 h and 72 h.
