AIM： To understand the local pathophysiological alterations and gene ontology-based functional classification of colonic biopsies into inflammatory and neoplastic diseases. METHODS： Total RNA was extracted from froz...AIM： To understand the local pathophysiological alterations and gene ontology-based functional classification of colonic biopsies into inflammatory and neoplastic diseases. METHODS： Total RNA was extracted from frozen biopsies and amplified by T7-method. Expression profile was evaluated by Atlas Glass 1K microarrays. After microarray quality control, applicable data were available from 10 adenomas, 6 colorectal adenocarcinomas （CRCs）, and 6 inflammatory bowel diseases （IBDs）. Multivariate statistical and cell functional analyses were performed. Real-time RT-PCR and immunohistochemistry were used for validation. RESULTS： Discriminant analysis of selected genes, could correctly reclassify all 22 samples using 4 parameters （heat shock transcription factor-l, bystin-like, calgranulin-A, TRAIL receptor 3）. IBD samples were characterized by overregulated chemokine （C-X-C motif） ligand 13, replication protein A1, E74-1ike factor 2 and downregu- fated TNF receptor-associated factor 6, BCL2-interacting killer genes. In adenomas upregulation of TNF receptorassociated factor 6, replication protein A1, E74-1ike factor 2 and underexpression of BCL2-associated X protein, calgranulin-A genes were found. CRC cases had significantly increased epidermal growth factor receptor, topoisomerase-1, v-jun, TNF receptor-associated factor 6 and TRAIL receptor 3, and decreased RAD51 and RAD52 DNA repair gene, protein phosphatase-2A and BCL2-interacting killer mRNA levels. Epidermal growth factor receptor RT-PCR and immunohistochemistry, topoisomerase-1 RT-PCR confirmed the chip results .CONCLUSION： Different histological alterations can be reclassified by functional, multivariate analysis using cDNA microarrays. Further studies with expanded sample number are needed for subclassification of pathological alterations.展开更多
A novel high-throughput system, called the stacked slice-gel system for separation and reactions （4SR）, was developed for the analysis of DNA/RNA and protein/peptide. The system provides a novel three-dimensional ge...A novel high-throughput system, called the stacked slice-gel system for separation and reactions （4SR）, was developed for the analysis of DNA/RNA and protein/peptide. The system provides a novel three-dimensional gel electrophoresis approach that exploits the property of stacked slice gels. It allows multiple samples simultaneously to react as well as to be separated, offering a two-dimensional （m×n） sample loading system. For this purpose, high-throughput multi-micro vessels （MMVs） containing variable numbers of wells （100 wells in this paper） have been used, which are made of 25 mm square-size polyacrylamide gels. Furthermore, after electrophoretic separation, a slice gel containing a desired sample can be easily removed and proceeded to the next step. Different biological reactions as well as successive separation of products were effectively carried out dealing with DNA/RNA and protein/peptide. It shows that this system has a diversity of potentials to be developed.展开更多
文摘AIM： To understand the local pathophysiological alterations and gene ontology-based functional classification of colonic biopsies into inflammatory and neoplastic diseases. METHODS： Total RNA was extracted from frozen biopsies and amplified by T7-method. Expression profile was evaluated by Atlas Glass 1K microarrays. After microarray quality control, applicable data were available from 10 adenomas, 6 colorectal adenocarcinomas （CRCs）, and 6 inflammatory bowel diseases （IBDs）. Multivariate statistical and cell functional analyses were performed. Real-time RT-PCR and immunohistochemistry were used for validation. RESULTS： Discriminant analysis of selected genes, could correctly reclassify all 22 samples using 4 parameters （heat shock transcription factor-l, bystin-like, calgranulin-A, TRAIL receptor 3）. IBD samples were characterized by overregulated chemokine （C-X-C motif） ligand 13, replication protein A1, E74-1ike factor 2 and downregu- fated TNF receptor-associated factor 6, BCL2-interacting killer genes. In adenomas upregulation of TNF receptorassociated factor 6, replication protein A1, E74-1ike factor 2 and underexpression of BCL2-associated X protein, calgranulin-A genes were found. CRC cases had significantly increased epidermal growth factor receptor, topoisomerase-1, v-jun, TNF receptor-associated factor 6 and TRAIL receptor 3, and decreased RAD51 and RAD52 DNA repair gene, protein phosphatase-2A and BCL2-interacting killer mRNA levels. Epidermal growth factor receptor RT-PCR and immunohistochemistry, topoisomerase-1 RT-PCR confirmed the chip results .CONCLUSION： Different histological alterations can be reclassified by functional, multivariate analysis using cDNA microarrays. Further studies with expanded sample number are needed for subclassification of pathological alterations.
文摘A novel high-throughput system, called the stacked slice-gel system for separation and reactions （4SR）, was developed for the analysis of DNA/RNA and protein/peptide. The system provides a novel three-dimensional gel electrophoresis approach that exploits the property of stacked slice gels. It allows multiple samples simultaneously to react as well as to be separated, offering a two-dimensional （m×n） sample loading system. For this purpose, high-throughput multi-micro vessels （MMVs） containing variable numbers of wells （100 wells in this paper） have been used, which are made of 25 mm square-size polyacrylamide gels. Furthermore, after electrophoretic separation, a slice gel containing a desired sample can be easily removed and proceeded to the next step. Different biological reactions as well as successive separation of products were effectively carried out dealing with DNA/RNA and protein/peptide. It shows that this system has a diversity of potentials to be developed.
