Screening the human pancreatic cancer-associated genes by cDNA microarray

Objective To screen the human pancreatic cancer-associated genes by cDNA microarray.Methods The PCR products of 14 000 genes were spotted onto a chemical-material-coated-glass plates in array.DNAs were fixed onto the glass plate after series of treatments.The total RNAs were isolated from 4 specimens of the pancreatic cancer and the normal tumor-surrounding pancreatic tissues.Both equal quantity RNAs were reversely transcribed to cDNAs and labeled with the fluorescent Cy5-dCTP and Cy3-dCTP to prepare the hybridization probes.The mixed probes were hybridized to the cDNA microarray.After high-stringent washing,the fluorescent signals were scanned by ScanArray 4000 scanner.The values of Cy5-dCTP and Cy3-dCTP on each spot were analyzed and calculated by GenePix Pro 3.0 software.Results By applying this cDNA microarray,189 differentially expressed genes in pancreatic cancer,whose ratios of Cy5/Cy3 were higher than 2.0 or lower than 0.5,were screened out among the 14 000 target genes,comprising 101 known genes and 88 novel gene.Among the known genes,upregulated and downregulated genes were 50 and 51 respectively,including oncogenes and tumor suppression genes,cell cycle related genes,cell skeleton and cinetc-protein related genes,cell apoptosis related genes,DNA synthesis related genes,DNA damage and repair-related genes,transcription factors,receptor related genes,immune related genes,signal transduction related genes,protein translation and synthesis related genes,metabolism related genes.Conclusion The analysis of gene expression profile of tumor based on cDNA microarray can realize high-throughput screening of the genes associated with the pancreatic cancer,and it can help to rapidly explore the genes function.Further analysis of the obtained genes will help to understand the molecular mechanism of the pancreatic cancer.