Project description:TGFb and/or crizotinib were treated on A549 cell line to understand molecular mechanisms of action of crizotinib in TGF signaling pathway in lung cancer through gene expression profile data.
Project description:Crizotinib is a first-line targeted therapy against EML4-ALK fusion in non-small cell lung cancer (NSCLC), but the increased resistance limits its clinical application. However, the mechanisms underlying such resistance are largely unexplored. The present study identified ALK protein overexpression in crizotinib-resistant H3122CR cells through immunoblotting. Then, liquid chromatography tandem mass spectrometry-based metabolomics was adopted to investigate metabolic responses. Crizotinib resistance was associated with the reduction of the total lipid content and the increase of organic acid, featuring the upregulation of lipid metabolism. Fatty acid β-oxidation pathway reprogrammed crizotinib-sensitive metabolome to crizotinib-resistant metabolome, resulting in crizotinib resistance. These findings reveal a previously unknown mechanism of crizotinib resistance and suggest promising applications of this approach in prognostic prediction and tailored therapeutics of NSCLC patients.
Project description:To investigate the difference of miRNA expression between lung cancer cell A549 and its DDP-resistant cell strain A549/DDP, we have employed miRNA microarray expression to discover the difference expression of miRNAs of A549 cells and A549/DDP. We conducted RT-qPCR to examine the expression levels of top differential expressed miRNAs, namely, miR-197-5p, miR-4443, miR-642a-3p, miR-27b-3p and miR-100-5p, confirming low variability between two methods. The A549/DDP was established from A549 in our laboratory, by exposing A549 to gradually increasing DDP concentrations, until the final concentration at 1μg/ml. To avoid the influence of drug to the A549/DDP cells, they were cultured in a drug-free medium for at least two weeks before gene expression analysis. miRNA expression of A549 and A549/DDP was then analzyed.
Project description:To investigate the effect of shikonin on the proliferation and migration of lung cancer cells, we used shikonin treated with A549 and H1299 cells We then performed gene expression profiling using RNA-seq data from A549 cells.
Project description:The anaplastic lymphoma kinase (ALK) gene fusion functions as a potent oncogene in lung cancer. The development of ALK tyrosine kinase inhibitors has improved patient outcomes. However, cancer progression commonly occurs even after treatment. Chemoresistance arises mostly by the acquisition of gatekeeper mutations in ALK as well as by off-target bypass mechanisms arising during chemotherapy. To investigate these bypass mechanisms, we studied crizotinib resistance in ALK-rearranged lung cancer by using 2D and 3D cell line models and longitudinal human biopsy samples. We found that crizotinib-resistant H2228 cells exhibited increased tumor cell clustering and spheroid formation, with elevated E-cadherin levels. Notably, depletion of E-cadherin profoundly disrupted spheroid formation and restored drug sensitivity. Transcriptomic profiling revealed the upregulation of tissue morphology-related genes; in particular, EpCAM was highly expressed in the protrusive regions of spheroids. Remarkably, longitudinal biopsy analysis also demonstrated enhanced expression of E-cadherin and EpCAM in patients with ALK-rearranged lung cancer during chemotherapy. We propose that off-target resistance is mediated by the upregulation of morphogenic genes, which drive cell clustering and spheroid formation during tumor evolution. To overcome chemoresistance, it is crucial to monitor tumor evolution during treatment. Tracking the collective migration of cells may present a novel therapeutic target.
Project description:To investigate the function SF3B4 during the progression of lung cancer, A549 cells with either SF3B4 siRNA or scramble siRNA were used for RNA-seq.
Project description:To study the effect of rs1663689 on lung cancer cells, we generated A549(C/C) from A549(T/T) using CRISPR/Cas9 technology and performed RNA-seq.
Project description:Resveratrol, a natural phytoestrogen found in red wine and a variety of plants, is reported to have protective effects against lung cancer, however there is very little work directed towards the understanding of the mechanism of action of resveratrol in lung cancer. In this study we used an experimental approach to understand the biological activity and molecular mechanisms of resveratrol in A549 lung cancer cells. Gene expression profiles were compiled using an oligonucleotide microarray to determine altered expression levels in resveratrol treated cells. Keywords: Genetic modification of A549 cells in response to resveratrol
Project description:To investigate the difference of miRNA expression between lung cancer cell A549 and its DDP-resistant cell strain A549/DDP, we have employed miRNA microarray expression to discover the difference expression of miRNAs of A549 cells and A549/DDP. We conducted RT-qPCR to examine the expression levels of top differential expressed miRNAs, namely, miR-197-5p, miR-4443, miR-642a-3p, miR-27b-3p and miR-100-5p, confirming low variability between two methods.