Global expression data from HCT-8 human colorectal cancer (CRC) wild type cells (HCT8/WT) and its 5-FU-induced resistant cell line (HCT8/5-FU)
ABSTRACT: Complete growth medium RPMI-1640 which contains 10% FCS, 1% Penicillin-Streptomycin Solution was used for cell culture. Both cell lines were maintained at 37 °C in a humidified atmosphere containing 5% CO2. MTT assay was performed to determine the 5 ‑ FU resistance in both of HCT-8/WT and HCT-8/5-FU cell line. Overall design: CRC wild type cell (HCT8/WT) and its 5-FU-induced resistant cell line (HCT8/5-FU) , both of which were treated with 5-fluorouracil (5-FU) for 0, 24 and 48 hours, respectively. Three biological replicates of each condition were used for the microarray experiment.
INSTRUMENT(S): [PrimeView] Affymetrix Human Gene Expression Array
A drug-induced resistant cancer cell is different from its parent cell in transcriptional response to drug treatment. The distinct transcriptional response pattern of a drug-induced resistant cancer cell to drug treatment might be introduced by acquired DNA methylation aberration in the cell exposing to sustained drug stimulation. In this study, we performed both transcriptional and DNA methylation profiles of the HCT-8 wild-type cells (HCT-8/WT) for human colorectal cancer (CRC) and the 5-fluor ...[more]
Project description:Complete growth medium RPMI-1640 which contains 10% FCS, 1% Penicillin-Streptomycin Solution was used for cell culture. Both cell lines were maintained at 37 °C in a humidified atmosphere containing 5% CO2. MTT assay was performed to determine the 5 ‑ FU resistance in both of HCT-8/WT and HCT-8/5-FU cell line. Overall design: CRC wild type cell (HCT8/WT) and its 5-FU-induced resistant cell line (HCT8/5-FU) , both of which were treated with 5-fluorouracil (5-FU) for 0 and 72 hours, respectively. Three biological replicates of each condition were hybridised to the Illumina Infinium 450k Human Methylation Beadchip.
Project description:We analyzed the differentially regulated genes in 5-fluorouracil-resnstant human colon cancer cells to discover novel biomarkers involved in 5-FU resistance in colorectal cancer. Overall design: The 5-FU resistant colorectal cancer cells were generated using HCT 116 human colon cancer cell line by treatment of cells with increasing concentrations of 5-FU more than 6 months. The basal gene expression profile in parental and 5-FU-resistant cells were analyzed and compared.
Project description:In pancreatic cancer the survival rate is low, as the available treatment options usually only extend survival and seldom produce a cure. Drug resistance and disease reoccurrence is the typical reason for death after cancer diagnosis. 5-Fluorouracil (5-FU) is the main chemostatic used in first line therapy. However the majority of the tumors become resistant to treatment. To investigate acquired 5-FU resistance in pancreatic adenocarcinoma, we established chemoresistant monoclonal cell lines from the Panc03.27 cell line by long-term exposure to 5-FU. In addition to increased expression of markers associated with multidrug resistance, the 5-FU resistant clones showed alterations typical of the process of epithelial-to-mesenchymal transition (EMT), including upregulation of mesenchymal markers and increased invasiveness. Microarray analysis revealed the L1CAM pathway as one of the most upregulated pathways in the chemoresistant clones, which was confirmed on RNA and protein levels. Expression of the adhesion molecule L1CAM is associated with a chemoresistant and migratory phenotype of pancreatic cancer. Using esiRNA targeting L1CAM, or by blocking the extracellular part of L1CAM with monoclonal antibodies, we discovered that the increased invasiveness observed in the chemoresistant cells depends on L1CAM. Using esiRNA targeting β-catenin and/or Slug, we discovered that L1CAM expression depends on Slug rather than β-catenin in the 5-FU resistant cells. We demonstrate a functional link between Slug and the expression level of L1CAM in pancreatic cancer cells having undergone EMT following long-term exposure to 5-FU. Our findings provide further insight into the molecular mechanisms leading to a chemoresistant and migratory phenotype in pancreatic cancer cells and indicate the importance of Slug-induced L1CAM in refractory pancreatic cancer. Examination of expression of 5-Fluorouracil (5-FU) Panc03.27 cell line resistant clone versus expression of 5-FU sensitive clones (NT) in 4 replicates per cell lines
Project description:The fluoropyrimidine 5-fluorouracil (5-FU) is an antimetabolite that exerts its anticancer effects through inhibition of DNA synthesis and replication by inhibiting thymidylate synthase, and by incorporation of its metabolites into RNA and DNA. 5-FU is used in most of the currently applied regimens for the treatment of patients with cancers of the gastrointestinal tract, breast, head, neck, and belongs to the most consumed cytostatic drugs. After application, 5-FU is via hospital and municipal waste water effluents released into the aquatic environment, where it can harm non-target organisms. The aim of this study was to evaluate changes in gene transcription in liver of adult F1 zebrafish (Danio rerio) continuously exposed to 5-FU at environmentaly relevant concentrations of 0.01 µg/L and 1 µg/L. One color design, control and treatment in two concentrations; 6 pools of 3 individuals per treatment
Project description:TE-5 and 5-FU resistant subline TE-5R Overall design: TE-5 cells were treated with continuous and step-wise concentrations of 5-FU (1, 2, 5, and 10 μM), and consequently 5-FU-resistant ESCC cells were established and named as TE-5R cells.
Project description:Cancer stem cells (CSCs) residing in colorectal cancer tissues have tumorigenic capacity and contribute to chemotherapeutic resistance and disease relapse. It is well known that the survival of colorectal CSCs after 5-Fluorouracil (5-FU)-based therapy leads to cancer recurrence. Thus CSCs represent a promising drug target. Here, we designed and synthesized novel hybrid molecules linking 5-FU with the plant-derived compound thymoquinone (TQ) and tested the potential of individual compounds and their combination to eliminate colorectal CSCs. SARB hybrid showed augmented cytotoxicity against colorectal cancer cells. Data obtained via Nanostring nCounter Gene Expression Assay (Pan Cancer Pathway Panel). Nanostring analysis revealed a unique signature of dysregulated gene expression in response to the combination of TQ and 5-FU (Combi) and SARB treatment. Importantly, two principle stem cell regulatory pathways WNT/ß-Catenin and PI3K/AKT were found to be downregulated after hybrid treatment. Overall design: In this study three biological replicates of each treatment and of the untreated control cells were included. HCT116 cells were treated for 24 h with the individual compounds, the combination of them and with the hybrid compound. We aimed to reveal differences in gene expression between treated and untreated cells as well as the differences on gene expression among different treatments.