Distinct DNA methylation profiles between adenocarcinoma and squamous cell carcinoma of human uterine cervix
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
SUBMITTER: Je-Ho Lee
PROVIDER: E-GEOD-15113 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:A microarray-based strategy using digestion with the methylation-sensitive restriction enzyme HpaII, ligation, and PCR, was performed to assess the DNA methylation status of promoter regions in Six human cervical adenocarcinomas (ACA) and four squamous cell carcinomas (SCC). Two array platforms were used; an array of 11,994 ~1.5kb PCR products from 10,445 promoter regions, and an array of 355,264 oligonucleotides for 18,212 HpaII fragments in 12,617 promoter regions. Loci near 21 genes showed significant differences between ACA and SCC. Real time PCR-based validation was performed on 13 loci using other nearby candidate methylation targets in the same promoter. Methylation patterns of eleven of these 13 linked loci concurred with the microarray results. Four loci were further studied using tissues from additional patients. Hyper-methylation of loci in PAK6 and NOGOR most strongly correlated with adenocarcinoma.
2009-03-18 | GSE15113 | GEO
Project description:Two prostate immortalized cell lines, 267B1, RWPE-1, Ml-csv40, and five prostate cancer cell lines, PC3, PC3M, PC3M-Pro4, PC3M-LN4 and LNCaP are included in this experiment. Their methylation profile are detected using human promoter array (v11). mRNA expression level are profiled with Affymetrix U133A Arrays for 267B1 and PC3M. Keywords: other
2010-06-09 | E-GEOD-951 | biostudies-arrayexpress
Project description:OsMKK4 is a rice MAPKK and immediately activated by treatment with chitin elicitor, a fungal MAMP. OsMKK4DD is a constitutively active mutants of OsMKK4. We identified OsMKK4 regulated genes using a conditional gain-of-function transgenic system and rice 44K oligoarray. We used transgenic rice expressing OsMKK4 in wild-type and constitutive active forms under the dexamethazone (DEX)-inducible promoter. Callus of the DEX inducible OsMKK4DD or OsMKK4WT line was compared with vector control line using two-color method with 3 biological replicates.
2010-07-30 | E-GEOD-15613 | biostudies-arrayexpress
Project description:Detection of copy number dependent gene expression profiles in epithelial tumours has lead to the discovery of genes significant in diagnosis, prognosis and of therapeutic value. We sought to apply the technologies of gene expression profiling and copy number aberration detection by high density oligonucleotide microarray analysis to detect genes with expression profiles regulated by copy number. We surveyed copy number aberrations in 27 squamous cell carcinomas tissues and ten cervical carcinoma cell lines by Agilent 244k aCGH, and in a companion study of 77 tissues we obtained transcriptional profiles of a broader ranger of histologies. Briefly, for normal (NAD) and squamous intraepithelial lesions (SILs), DNA and RNA were extracted from approximately 5 - 25 x 5 mm thin sections of fresh frozen cervical tissue, respectively. Epithelial were microdissected, pooled and processed as follows. For RNA extraction, tissue was collected in Trizol (Invitrogen, UK) and subject to homogenization (Polytron), total RNA was recovered following standard processes, and quality assessed by BioAnalyzer (Agilent). DNA was recovered following Proteinase K digestion, phenol chloroform extraction, and ethanol precipitation. Contaminating RNA was removed by RNAse A treatment, and a second round of phenol chloroform extraction and ethanol precipitation. For SCCs, RNA and DNA was extracted from whole tumour (>85%), following similar strategies as previously described. This submission describes the copy number aberrations of 27 cervical SCC tissues and 10 common cervical SCC cell lines
2011-03-05 | E-GEOD-27673 | biostudies-arrayexpress
Project description:The neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP) is considered to be a potential therapeutic agent for prevention of cerebral ischemia. Ischemia is a most common cause of death after heart attack and cancer causing major negative social and economic consequences. - This study was designed to investigate the effect of PACAP38 injection intracerebroventrically in a mouse model of permanent middle cerebral artery occlusion (PMCAO) along with corresponding sham control that used 0.9% saline injection. Ischemic and non-ischemic brain tissues were sampled at 6 and 24 hours post-treatment. - Following behavioral analyses to confirm whether the ischemia has occurred, we investigated the genome-wide changes in gene and protein expresison using DNA microarray chip (4x44K, Agilent) and one-/two-dimensional gel electrophoresis (1-/2-DGE) coupled with matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS), respectively. - Our results revealed numerous changes in the transcriptome of ischemic (ipsilateral) hemisphere treated with PACAP38 over the saline-injected SHAM control hemisphere. In parallel, 2-DGE analysis revealed a highly expressed protein spot in the ischemic region that was identified as dihydropyrimidinase-related protein 2 (DPYL2). The DPYL2, also known as Crmp2, is a marker for the activated neuronal defense mechanisms, and interestingly, PACAP strongly suppresed this induction. - This study provides the first inventory of PACAP influenced gene and protein targets in ischemic hemisphere, and provides new targets for thereaupetic interventions. The PMCAO model mice were generated as described previously. Briefly, the mice were anesthetized with 4% sevoflurane (induction) and 2% sevoflurane (maintenance) in a 30% O2 and 70% N2O gas mixture via a face mask. An incision was then made in the cervical skin followed by opening of the salivary gland, and the right common carotid artery was visualized. A midline cervical incision was made to expose the external carotid artery. The intraluminal filament technique was used, to generate the PMCAO model. - The PACAP38 (1 M-BM-5l containing 1 pmol) or 1 M-BM-5l of saline (0.9% NaCl) was injected intracerebroventrically, immediately after PMCAO. PACAP38 (Peptide Institute Inc., Osaka, Japan; supplier temperature was -20M-BM-:C and was stored at -80M-BM-:C) was dissolved in, and diluted M-CM-^W10 times with 0.9% NaCl just before use. - In the sham control animals, following exposure of the external carotid artery, the wound was sutured. The PACAP38 or saline was injected intracerebroventrically in the same concentrations as above. After injections, the animals were returned to their cages. - A total of eight groups were prepared: four groups of three, seven, four and five mice in the PMCAO plus PACAP38 and PMCAO plus saline cohorts at 6 and 24 h after operation, respectively, and five mice each in the control (sham) plus PACAP and saline groups at 6 and 24 h after operation, respectively. - We used three mice each in PMCAO groups that exhibited neurological grades G1 and G2 and three mice each at random in sham groups for the subsequent downstream analysis. Six or 24 hours post-injection of PACAP38 or saline, the mice were removed from their cages, decapitated, and their brains carefully removed on ice. The left (contralateral) and right (ipsilateral; ischemic) hemispheres were dissected and placed in 2 mL Eppendorf tubes, which were then quickly immersed in liquid nitrogen before being stored in -80M-BM-:C prior to further analysis. - For analysis of gene components, the tissues were ground to a very fine powder with liquid nitrogen, used for total RNA extraction, quantity and quality check, followed by cDNA synthesis, and RT-PCR. A mouse 4 x 44K whole genome oligo DNA microarray chip (G4122F, Agilent Technologies, Palo Alto, CA, USA) was used for global gene expression analysis using the ipsilateral (ischemic) hemisphere. Briefly, total RNA (900 ng; 300 ng each replicate pooled together) was labeled with either Cy3 or Cy5 dye using an Agilent Low RNA Input Fluorescent Linear Amplification Kit (Agilent). Fluorescently labeled targets of control (sham) as well as treated (PMCAO) samples with PACAP38 or without PACAP38 (saline) were hybridized to the same microarray slide with 60-mer probes. - Here, we compared the PMCAO plus PACAP38 injected mice to PMCAO plus saline, i.e. the ipsilateral brain region of the PMCAO mice was compared with the same right hemisphere of the control mice. Similarly, Sham control plus PACAP38 injected mice to Sham control plus saline were analyzed to identify only the PACAP38 influenced genes. A flip labeling (dye-swap or reverse labeling with Cy3 and Cy5 dyes) procedure was followed to nullify the dye bias associated with unequal incorporation of the two Cy dyes into cDNA.
2012-04-30 | E-GEOD-37565 | biostudies-arrayexpress
Project description:High ozone (O3) concentration causes serious damages in plant productivity. Climate models forecast that ground O3 level in the future will reach phytotoxic range, resulting in crop yield losses. With an ultimate goal to screen molecular factors to minimize losses of crop production by the rise of O3 level, we have started an investigation on effects of O3 on rice using rice DNA chip. Herein, we have utilized the samples of dry mature rice seeds harvested in an ozone-sensitive rice cultivar (Oryza sativa L. indica cv. Takanari) and a tolerant cultivar (Oryza sativa L. japonica cv. Koshihikari) which were fumigated with ambient air (mean O3: 32.7 ppb) in small open-top chambers (OTCs). First, we extracted total RNA from dry mature rice seeds of Takanari and Koshihikari using a modified protocol based on cethyltrimethylammonium bromide extraction buffer and phenol-chloroform-isoamylalcohol treatment. Furthermore, to perform microarray analysis using the Agilent 4x44 rice DNA Chip and the dye-swap method, we designed a balanced block design comparing seeds in an ambient air-fumigated rice cultivar and those in a filtered air-fumigated rice cultivar. Direct comparison of Koshihikari and Takanari O3 transcriptomes in seeds of rice plants fumigated with ambient O3 in OTCs successfully showed that genes encoding proteins involved in jasmonic acid, GABA biosynthesis, cell wall and membrane modification, starch mobilization, and secondary metabolite biosynthesis are differently regulated in an O3-sensitive cv. Takanari and a tolerant cv. Koshihikari.
2012-04-30 | E-GEOD-37650 | biostudies-arrayexpress
Project description:Background: N-Methyl-D-aspartate receptors (NMDAr), widely located around the central nervous system, are known to be involved in behavioral disorders. Dizocilpine (commonly referred to as MK-801) is a well known non-competitive NMDAr antagonist. Methods: We treated rats with intraperitoneal injection [0.08 (low-dose) and 0.16 (high-dose) mg/kg] of MK-801. In one experiment, 40 min after NaCl (vehicle control) and MK-801 (0.08 mg/kg) injection, electrocorticogram (ECoG) signals were analyzed. In the second experiment, 40 min post-injection, the whole brain of each animal was rapidly removed and separated into amyglada, cerebral cortex, hippocampus, hypothalamus, midbrain and ventral striatum) on ice, followed by analysis using a 4x44K DNA microarray chip. Results: Spectral analysis revealed that a single subcutaneous injection of MK-801 significantly and selectively augmented the power of spontaneous gamma and higher-frequency oscillations. The results from DNA microarray analysis of 4400 genes showed the largest number (up- and down- regulations) of gene expressions in the cerebral cortex (378), midbrain (376), hippocampus (375), ventral striatum (353), amygdala (301), and hypothalamus (201) under low-dose of MK-801. Under high-dose, ventral striatum (811) showed the largest number of gene expression changes. These genes represented... Conclusions: Our results reveal that MK-801 triggered i) an increase in the power of gamma oscillations, and ii) simultaneously caused very early changes in gene expressions in the rat brain, representing a first such inventory of gene expression profiles in brain after acute MK-801 treatment. Nine male 10-weeks-old Wistar rats (300-350 g BW) were housed in acrylic cages (3/cage) at 24ºC and given access to tap water and laboratory chow ad libitum. The rats were divided into two groups, and each group rats received i.p. injection of 0.08 (low-dose) and 0.16 (high-dose) mg/kg of MK-801, respectively. Three rats were treated with saline as sham (vehicle control group) using the same method. After 40 min post-injection, the whole brain of each animal was rapidly removed and put on ice, and brain regions were separated according to the method of Glowinski and Iversen (1996), with minor modifications (Hirano et al., 2007). Each brain region was placed in a 2 mL Eppendorf tube, quickly immersed in liquid nitrogen before being stored in -80ºC prior to further analysis. Each sample was immediately weighed, flash-frozen in liquid nitrogen and stored at -80ºC prior to further analysis. A rat 4 x 44K whole genome oligo DNA microarray chip (G4131F, Agilent Technologies, Palo Alto, CA, USA) was used for global gene expression analysis. The effects of MK-801 were examined in the 6 brain reagion, Ventral striatum, Cerebral cortex, Midbrain, Amygdala, Hippocampus, and Hypothalamus.
2012-04-24 | E-GEOD-37413 | biostudies-arrayexpress
Project description:A combined approach of evaluating ozone (O3)-caused foliar injury symptom and global gene expression profiling was used to identify potential genes associated with severity of injury on leaves of O3 (200 ppb)-fumigated (1, 12, 24, 48, and 72 h) two-week-old rice (cv. Nipponbare) seedling along with appropriate control. Foliar injuries were evaluated up to 72 h using both qualitative visual scale and quantitative RGB (red-green-blue) image analysis methods. The (R-G)/(R+G) was found as the optimal quantitative RGB parameter to assess the foliar injury. Large-scale transcript profiling of leaves identified 270 genes linked with foliar injury. Of these genes, the expression levels of 139 genes showed significant differences (P < 0.05) between leaves without and with injury symptoms. When a rigorous correlation test was applied on these genes for their expression changes and relative (R-G)/(R+G) parameters, the expression of 93 genes were found to increase with increased foliar severity up to 72 h, showing a positive, tight correlation between a subset of gene expression and commonly observed O3-triggerred symptom of foliar injury with correlation coefficient below -0.80. Of 93 genes, genes involved in metabolism (29%) formed a major functional category. Reconstruction of the metabolic networks with identified metabolic genes provided insight into the cellular responses such as photorespiration, biosynthesis of secondary metabolites, and detoxification. The O3 effect on these cellular responses has been previously reported based on physiological and biochemical studies, validating our approach used in this study to globally identify O3-responsive biomarkers tightly linked with foliar injury symptom. This study provides evidence for the presence of large number of genes associated with the foliar injury symptom than thought before, and could serve as a resource of potential biomarkers to study mechanisms of visible injury development by O3. Comparison between healthy rice seedling leaves and ozone treated (for 48 h and 72 h) rice seedling leaves was performed. Three biological replicates (5 leaves in each replicate; pooled) were used, and dye-swaped.
2011-12-10 | E-GEOD-34256 | biostudies-arrayexpress
Project description:There is increasing evidence for tissue stem cells as potential source for cancers. However, the cellular and molecular mechanisms at the origin of this transformation remain elusive. Here we show that Delta-like1 (Dll1) mutation induces the oncogenic transformation of neural stem cell–derived neurospheres into oncogenic spheres capable of generating histological and molecular human glioblastoma (GBM)-like tumors upon subcutaneous and intracranial transplantations into nude mice. Serial transplantation assays suggest that Dll1 spheres tumorigenicity results from the oncogenic transformation of normal neural stem cell into GBM-producing stem cell. Compared differentiation of Dll1 vs. WT spheres shows the presence in Dll1 spheres of a subpopulation of cells which although fated to a glial lineage fail to differentiate terminally, in keeping with the astrocytic promoting function of Delta-Notch. This population of astrocyte progenitors proliferates actively and fails to down-regulate EGFR phosphorylation. These studies suggest that Dll1-induced tumorigenesis is restricted to the astrocytic lineage thereby providing a model for human GBM as well as an explanation for the duality of Notch signaling acting either as an oncogene or a tumor suppressor in stem/progenitor cells derived tumors, depending on the lineage commitment. Comparison between control (WT) and mutants (Dll1-3 and Dll1-5) cells was performed. Two biological replicates (n=3 each replicate) were used, and each replicate (n=3, total RNA pooled) was dye-swaped.
2011-09-23 | E-GEOD-32293 | biostudies-arrayexpress
Project description:The ‘O3-responsive transcriptome’ behavior in the panicles and grains of rice plant was studied individually through high-throughput oligo-DNA microarray technique. Obtained results showed that O3 differentially regulated 620 and 130 genes in the panicles and grains separately, by at least two-fold changes. However, only five genes were found to be common in both the tissues, suggesting towards the tissue specific O3-sensitivity in rice plants. Among the O3-responsive genes, 176 and 444 genes were up- and down-regulated in rice panicle; whereas, 24 and 106 genes in rice grain, respectively. Further mapping onto various regulatory events revealed that, the majority of differentially expressed genes were mainly involved in signaling, hormonal, cell wall, transcription, proteolysis, and defense events. Many previously unknown O3-responsive novel genes were identified, including the brassinosteroid insensitive-1 receptor kinase, wall-associated kinase like receptor, calcium-dependent protein kinases, phosphatidylinositol kinases, G-protein components, ethylene insensitive-3, cellulose synthases, pectatelyase, etc. Inventory of 745 O3-responsive genes and their mapping will surely expand our knowledge on novel regulatory processes in both panicles and grains of rice; and, serve as a resource towards the designing of rice crops for future high-O3 world. Comparison between healthy rice plant panicles and ozone treated plant panicles (for 8 h) and seed (grain) of healthy rice plants and of rice plants grown under ozone for their lifeftime was performed. Three biological replicates (panicle or seed; pooled) were used, and dye-swaped.
2012-04-24 | E-GEOD-37478 | biostudies-arrayexpress