Transcription profiling by array of human gastric cancer cells treated with suberoylanilide hydroxamic acid (SAHA)
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ABSTRACT: Gastric cancer cell line AGS was treated with suberoylanilide hydroxamic acid (SAHA) for 24 hours. Microarray analysis was done to explore the differentially expressed genes betweenSAHA treated and control cells.
Project description:To unravel antihypertensive drug-induced gene expression changes that are potentially related to the amelioration of end-organ damages, we performed in vivo phenotypic evaluation and transcriptomic analysis on the heart and the kidney, with administration of antihypertensive drugs to two inbred strains of (i.e., hypertensive and normotensive) rats. We chose to use six antihypertensive classes: enalapril (angiotensin converting enzyme inhibitor), candesartan (angiotensin receptor blocker), hydrochlorothiazide (diuretics), amlodipine (calcium-channel blocker), carvedilol (vasodilating beta-blocker) and hydralazine. In the tested rat strains, four of six drugs, including two renin-angiotensin system (RAS) inhibitors, were effective for BP lowering, whereas the remaining two drugs were not. Besides BP lowering, there appeared to be some inter-drug heterogeneity in phenotypic changes, such as suppressed body weight (Bw) gain and Bw-adjusted heart weight reduction. For the transcriptomic response, a considerable number of genes showed prominent mRNA expression changes either in a BP-dependent or BP-independent manner with substantial diversity between the target organs. Noticeable changes of mRNA expression were induced particularly by RAS blockade, e.g., for genes in the natriuretic peptide system (Nppb and Corin) in the heart and for those in the RAS/ kallikrein-kinin system (Ren and rat Klk1 paralogs) and those related to calcium ion binding (Calb1 and Slc8a1) in the kidney. This submission includes the kidney data.
Project description:AGS gastric cancer cell line was treated with the selective COX-2 inhibitor celecoxib. microRNA expression profile was analyzed between untreated cells and cells treated with celecoxib.
Project description:Gene expression profiling in human gastric cancer cells after 5-aza-2'-deoxycytidine (5-Aza-CdR) and 4-phenylbutyric acid (PBA) treatment was analyzed to investigate what genes are controlled by these chromatin modifying drug
Project description:Cyclic indole-3-carbinol (I3C) tetrameric derivative (CTet) is an anticancer molecule that has been shown to exert an antiproliferative activity in both MCF-7 and MDA-MB-231 breast cancer cell lines. To characterize the molecular mechanisms leading to the inhibition of the cell proliferation, gene expression analyses were conducted. MCF-7 and MDA-MB-231 breast cancer cells were plated in 6-wells culture plates at density of 150,000 cells/well and cultured overnight. Cellular treatments were conducted at 6uM and 12 uM concentration of CTet, or vehicle control, for 24 h. Cell survival was evaluated by trypan blue dye exclusion assay and, after washing in phosphate buffered saline (PBS), the cells were pelletted by centrifugation and stored at -20M-0C with 300M-5l of RNA-later solution (Sigma-aldrich). Total RNA was purified from treated and control cells using the RNeasy plus kit (Qiagen). Biotin-labeled cRNA was synthesized using the CodeLink iExpress Assay reagent kit (GE Healthcare), following the manufacturerM-^Rs protocols. Biotin-labeled cRNA obtained from each biological sample was fragmented and hybridized against three independent arrays (10 ug each) at 37M-0C for 22 h. After hybridization, the arrays were washed, stained with Cy5-streptavidin and scanned using a ScanArray GX scanner (Perkin Elmer), with a resolution of 5 um. The image files generated by the scanner were processed using the Codelink Expression Analysis software (GE Healthcare). Normalized data from the Codelink software package were analyzed with GeneSifter software (www.genesifter.net; Geospiza Inc., Seattle, WA) for statistical validation and data mining. Normalized data of the two experiments were subjected to analysis of variance (ANOVA) and 5% false discovery rate calculation (Benjamini and Hochberg). The cut-off parameters for differential gene expression were p =0.01 and fold change threshold =2.<br>
Project description:microRNA expression profiling in gastric cancer cells after 5-aza-2'-deoxycytidine (5-Aza-CdR) and 4-phenylbutyric acid (PBA) treatment was performed to investigate whether microRNA expression is controlled by these chromatin modifying drugs.
Project description:To determine the consequences of hypoxia-mediated HIPK2 clearance on gene expression, cells 293T were transfected with either the HIPK2 expression vector or the corresponding empty vector and then subjected to either hypoxia conditions (2% O2 for 6 h) or normoxia conditions. RNA total was extracted and later a study of the different gene expression under the four conditions detailed above was performed. The correct expression of HIPK2 was assessed by western blot in a small cell pool. The quality of RNA was quantified before the array study. <br><br>
Project description:Stimulation of the mouse hindlimb via the sciatic nerve was used to induce contractions for 4 hours to investigate acute muscle gene activation in a model of muscle phenotype conversion. Initial force production (1.6 + 0.1 g/g body weight) declined 45% within 10 min and was maintained for the remainder of the experiment. Force returned to initial levels upon completion of the study. An immediate-early growth response was present in the EDL (FOS, JUN, ATF3, MAFK) with a similar but attenuated pattern in the soleus. Transcript profiles showed decreased fast fiber specific mRNA (myosin heavy chains 2A, 2B; troponins T3, I; -tropomyosin, m-creatine kinase) and increased slow transcripts (myosin heavy chain slow/1, troponin C, tropomyosin 3) in the EDL. Histological analysis of the EDL revealed glycogen depletion without inflammatory cell infiltration or myofiber damage in stimulated vs. control muscles. Several fiber type specific transcription factors (EYA1, TEAD1, NFATc1 and c4, PPARG, PPARGC1 and β, BHLHB2) increased in the EDL along with transcription factors characteristic of embryogenesis (KLF4, SOX17, TCF15, PKNOX1, ELAV). No established in vivo satellite cell markers or the genes activated during our parallel studies of satellite cell proliferation in vitro (CYCLINS A2, B2, C, E1, MyoD) increased in the stimulated muscles. These data indicated that onset of fast to slow phenotype conversion occurred in the EDL within 4 hours of stimulation without satellite cell recruitment or muscle injury but was driven by phenotype specific transcription factors from resident fiber myonuclei including activation of nascent developmental transcriptional programs. Adult male Swiss Webster mice (30-35 g) were anesthetized, a bipolar electrode was implanted adjacent to the sciatic nerve and the hindlimb immobilized. The voltage-force relation was determined to establish supramaximal stimulation conditions and the length-tension relation was determined to set the resting length for maximum twitch tension. Contractions were induced by sciatic nerve stimulation (0.5 msec duration, 2-5 volts). The muscles were allowed to rest 15 minutes for full metabolic recovery at physiologic temperatures. Supramaximal stimulation was applied at a rate of 10 Hz for 4 hours. At the end of each experiment the soleus muscles were carefully dissected and flash frozen in liquid nitrogen for analysis of mRNA expression via microarray analysis. The contralateral, unstimulated Soleus provided a genetically matched, paired control for each specimen.
Project description:Evaluation of two commercial microarray platforms (Amersham CodeLink UniSet Human 10K I BioArray and Affymetrix GeneChip HG-U133A). Both platforms have been tested on gene expression profiling of MDA-MB-231 human metastatic breast cancer cells, cultured for 48 h in the absence (control) or presence (treated) of 32 µM resveratrol.
Project description:Measurement of cytokine secretion and gene expression changes from allergic and tolerant<br>patient lymphocytes stimulated with p-phenylenediamine and Bandrowski's base