ABSTRACT: Wnt signaling plays important roles in development and cancer. We found hiNOS is a wnt target gene. We tried to clarify the role of the so far iNOS in Wnt signaling in colorectal cancer. We performed SuperArray limited gene array analysis of SW480 cells with/without hiNOS overexpression. Oligo GEArray expression array systems (cat#OHS-043) SuperArray, Bethesda, MD) consisted of spotted cDNA fragments encoding 114 genes involved in and downstream of Wnt signaling. Control sequences (PUC18, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and ?-actin) were also included. These Superarrays were employed to compare wnt signaling genes expression between with iNOS and without iNOS. Total RNA was isolated by Trizol (Invitrogen) and processed for microarray hybridization following the manufacturer’s instructions. Arrays were visualized by autoradiography, and hybridization signals were scanned and analyzed for density in GEArray Expression Analysis Suite 2.0. The normalized value for each gene was calculated by dividing the value of each gene by the average value of the housekeeping genes GAPDH, and ?-actin. To obtain a gene profile, analysis was performed on three separate RNA samples isolated from different SW480 cells, with replicate assays conducted on each RNA preparation.
Project description:To determine gene expression profiles of neurogenesis, neural stem cell and notch signaling elements in the adult subventricular zone (SVZ) of the WT and CNP-hEGFR mouse, we used SuperArray limited gene array analysis. GEArrayTM expression array systems (cat#OMM404 SuperArray, Bethesda, MD) consisted of spotted cDNA fragments encoding 288 mouse genes related to neurogenesis and neural stem cell differentiation, as well as notch signaling elements. Control sequences (PUC18, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), peptidylpropyl isomerase A (Ppia), and β-actin) were also included. These microarrays were employed to compare SVZ gene expression between WT and CNP-hEGFR mouse SVZ. Total RNA was isolated by Trizol (Invitrogen) and processed for microarray hybridization following the manufacturer’s instructions. Arrays were visualized by autoradiography, and hybridization signals were scanned and analyzed for density in GEArray Expression Analysis Suite 2.0. The normalized value for each gene was calculated by dividing the value of each gene by the average value of the housekeeping genes GAPDH, Ppia, and β-actin. To obtain a gene profile, analysis was performed on three separate RNA samples isolated from different SVZs, with replicate assays conducted on each RNA preparation.
Project description:To determine expression profiles of cytokines and growth factor profiles in the subventricular zone (SVZ) after demyelination we performed gene array analysis. In SVZ after 4 days post lesin (dpl) LPC-injected tissue, demonstrated regulation of BMP pathway elements, including increased chordin, noggin, and ChorR, and decreased BMP4, compared to NaCl-injected tissue. GEArrayTM expression array systems (cat#OMM031 SuperArray, Bethesda, MD) consisted of spotted cDNA fragments encoding 113 mouse genes for neurotrophic signaling molecules involved in neuronal growth and differentiation, as well as regeneration and survival. Control sequences (PUC18, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), peptidylpropyl isomerase A (Ppia), and ?-actin) were also included. These microarrays were employed to compare SVZ gene expression 4 days after NaCl or LPC injection in corpus callosum. Total RNA was isolated by Trizol (Invitrogen) and processed for microarray hybridization following the manufacturer’s instructions. Arrays were visualized by autoradiography, and hybridization signals were scanned and analyzed for density in GEArray Expression Analysis Suite 2.0. The normalized value for each gene was calculated by dividing the value of each gene by the average value of the housekeeping genes GAPDH, Ppia, and ?-actin.
Project description:Accumulated research has suggested the importance of the adhesion molecules modulation as therapeutic approach for bronchial asthma. Adhesion molecules expression alteration contributes to the pathogenesis of asthma. In order to probe the relationship between expression imbalance of adhesion molecules and asthma pathogenesis, expression profiling of adhesion molecules was performed using cDNA microarray. The results showed that there were various adhesion molecules with abnormal expressions in peripheral blood leucocytes of asthma patients. RNA was extracted from leucocytes in peripheral blood of 4 normal adults and 6 asthma patients by using TRIzol Reagent. Microarray expression studies were performed using the GEArray Q Series Human Extracellular Matrix & Adhesion Molecules Gene Array (SABiosciences Corporation, USA). This microarray profiles the expression of 96 genes key to the functions of cell adhesion. A negative control (PUC18DNA and blank), and the housekeeping genes including β-actin, GAPDH, Cyclophilin A and ribose body protein L13a were spread on each chip.
Project description:[original title] Microarray analysis of DNA damage repair gene expression profiles in cervical cancer cells radioresistant to 252Cf neutron and X-rays. The aim of the study was to obtain stable radioresistant sub-lines from the human cervical cancer cell line HeLa by prolonged exposure to 252Cf neutron and X-rays. Radioresistance mechanisms were investigated in the resulting cells using SuperArray Oligo GEArray® Human DNA Damage Signaling Pathway Microarray.
Project description:To determine gene expression profiles of neurogenesis, neural stem cell and notch signaling elements in the adult subventricular zone (SVZ) of the WT and CNP-hEGFR mouse, we used SuperArray limited gene array analysis. GEArrayTM expression array systems (cat#OMM404 SuperArray, Bethesda, MD) consisted of spotted cDNA fragments encoding 288 mouse genes related to neurogenesis and neural stem cell differentiation, as well as notch signaling elements. Control sequences (PUC18, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), peptidylpropyl isomerase A (Ppia), and β-actin) were also included. These microarrays were employed to compare SVZ gene expression between WT and CNP-hEGFR mouse SVZ.
Project description:Colonizing commensal bacteria after birth are required for the proper development of the gastrointestinal tract. It is believed that bacterial colonization pattern in neonatal gut affects gut barrier function and immune system maturation. Studies on the development of faecal flora microbiota in infants on various formula feeds showed that the neonatal gut was first colonized with enterococci followed by other flora microbiota such as Bifidobacterium in breast feeding infants. Intriguingly, Bjorksten group Other studies showed that Bbabies who developed allergy were less often colonized with Enterococcus during the first month of life as compared to healthy infants. A lot of Many studies have been done on conducted to elucidate how bifidobacteria or lactobacilli, some of which are considered probiotic, regulate infant gut immunity. However, much fewer studies have been focused on enterococi. In our study, we demonstrate that E. faecalis, isolated from healthy newborns, suppress inflammatory responses activated in vivo and in vitro. We found E. faecalis attenuates proinflammatory cytokine secretions, especially IL-8, through JNK and p38 signaling pathways. This finding shed light on how the first colonizer, E.faecalis, regulate inflammatory responses in the host. Samples are analysed using web-based GEArray Expression Analysis Suite
Project description:We investigated the inflammatory gene profile of surgically-repaired lacerated muscles during the recovery phase and examined if it was influenced by the integrity/injury of the IM, intramuscular nerve using 3 lacerated rat muscle models: DN, where the IM-nerve was concomitantly cut; RN, where the IM-nerve was crushed but leaving an intact nerve sheath; and PN, a control where the IM-nerve was preserved intact. After 2, 4 and 8 weeks, the animals were sacrificed and their gastrocnemius muscles were removed and total RNA was extracted with Qiagen RNeasy Fibrous Tissue Mini Kit according to manufacturers instructions. Biotin-labelled cRNA probes were synthesized from total RNA by using a TrueLabeling-AMP Linear RNA amplification kit (SABiosciences Corp, Frederick, MD). The labeled cRNA probes were hybridized to oligonucleotide fragments spotted on the gene array membranes. Membranes were washed to remove any unincorporated probe and incubated with alkaline phosphatase-conjugated streptavidin (AP-streptavidin). Relative expression levels of specific genes were detected from signals generated by chemiluminescence from the alkaline phosphatase substrate, CDP-Star. The luminizing blots were used to expose X-ray films and quantified by spot densitometry with the aid of GEArray expression analysis suite (SABiosciences Corp, Frederick, MD). The abundance of each transcript was normalized to the normal un-operated muscle of the opposite limb, and to the housekeeping gene markers on each array (Aldoa, GAPdH and BAS2C). The results were presented as log2-fold expression with PN used as a control, to compare the two different types of nerve injuries (RN and DN).
Project description:The propensity for prostate cancer to metastasize to bone led us and others to propose that bidirectional interaction between prostate cancer cells and bone are critical for the preferential metastasis of PCa to bone. We previous identified a secreted isoform of ErbB3 (p45-sErbB3) from the bone marrow supernatants of patients with prostate cancer and bone metastasis. Immunohistochemical analysis of p45-sErbB3 expression in human specimens showed that p45-sErbB3 was highly expressed in metastatic prostate cancer cells in bone. Here we show that p45-sErbB3 stimulates calvarial bone to secrete factors that increase the invasiveness of prostate cancer cells in Boyden chamber invasion assay. We used gene array analysis to identify p45-sErbB3 regulated osteoblast genes that may enhance the invasiveness of PC-3 cells and found that p45-sErbB3 stimulated the expression of osteonectin, biglycan, and type I collagen in mouse calvaria. We further showed that recombinant osteonectin increases the invasiveness of PC-3 cells and osteonectin neutralizing antibody blocked p45-sErbB3 mediated invasiveness. These results suggest that p45-sErbB3 enhances the invasiveness of PC-3 cells is due, at least in part, to the stimulation of the secretion of osteonectin from bone. Thus, p45-sErbB3 mediates the bi-directional interaction between PCa cells and bone through osteonectin. Sample 1: non-treated calvaria RNA, Group 1 Sample 2: sErbB3 (100ng/ml) treated calvaria RNA, Group 2 Array: GE-supper array MM026 (total 96 genes, with 10 housekeeping genes) (service from SuperArray Bioscience Corporation: http://www.superarray.com
Project description:Metastasis is the major cause of cancer-related death in patients with colorectal cancer. Although inducible nitric oxide synthase (iNOS) is a crucial regulator of cancer development and progression, its roles in epithelial-mesenchymal transition (EMT) and the pathogenesis of metastatic colorectal cancer have not been fully investigated. Primary colorectal cancer and liver metastatic tissue specimens were analyzed showing 90% of liver metastatic colorectal cancer with reduced expressions of iNOS compared with 6% of primary colorectal cancer. The Cancer Genome Atlas database analyses via cBioPortal reveal that mRNA expression of iNOS negatively correlated with selected EMT markers in colorectal cancer in a cancer type-dependent manner. The transcriptomic profiling (RNA sequencing data) indicates that iNOS knockdown in SW480 colorectal cancer cells induced an EMT program with upregulated expression of selected stem-cell markers. iNOS knockdown did not alter E-cadherin mRNA expression but re-localized it from membrane to cytoplasm through iNOS-GATA4-Crb2-E-cadherin pathway. iNOS knockdown induced a change in cell morphology, and promoted cell invasion and migration in vitro, and metastasis in vivo.
Project description:Sera of experimental autoimmune encephalomyelitis (EAE) or control mice were collected at day 18 post immunization. Comprehensive analysis of cytokine levels were performed by using commercially available RayBio Mouse Cytokine Array 2 (RayBiotech, Inc.) according to manufacturer’s protocol 4 samples.There are two groups: vehicle control group and EAE group. Each group consists of 2 replications.