Study On Differentially Expressed Genes In Esophageal Cancer By Oligochip

Esophageal carcinoma is a common malignancy. Staging of the tumor has been proved to be associated with prognosis. So, it is most important for us to reveal the inbeing of esophageal carcinoma. Recently, with the development of molecular biology, it has been proved that the most common shifts of gene in the esophageal carcinoma are the activation of oncogene and inactivation of tumor suppressor gene, which lead to the occurrence and development of the cancer through taking part of the regulation of cell cycle, signal transfer, cell differentiation, apoptosis and so on. The gene chip technique, which is springing up recently, has shown great potential in the cancer research. The gene chip, in fact, is a high-density nucleic acid array, which fix up a large number of nucleic acid segments (probes) with special sequences on the slides, analyzing the sequences of samples of nucleic acid unkown by hybridization with probes. This technique can be used to sequences and quantitative analysis rapidly, largely and exactly.Purpose: To filtrate the genes associated with esophageal carcinoma and investigate the relationship between the genes and esophageal carcinoma by oligonucleotide chip technique. Materials and methods: 1.Material: one tissue sample supplied by the first affiliated hospital, medical college, Zhejianguniversity.Male ,58Y ,hospital number: 361149 , Pathologic diagnosis: medium-differentiatedsquamous cell carcinoma of esophagus.TNM type: T4N1M02.Methods:Pick up total RNA from esophggeal carcinoma, pericancerous tissues and normaltissue of esophagus, then retro-iranscribe cDNA and lable them with Cy3-duTP.According to literatures, 238 cancer-related genes were filtrated, whose sequenceswas found out from the Genebank, and were designed as probes by the software withthe length of 40 bp. These: oligonucleoti.de probes were arranged on the silylatedslides, composing a 12X24 matrix. The probes were hybridized with the gene chips(temperature: 40℃, Time: 3 hr). Fluorescence signals were scanned by laser scannerand farther analyzed by ImaGene4.0 software with parallel comparison among thesethree gene profiles.The difference of gene expression was judged when onefluorescence intensity, at least, among the three gene profiles should be larger than1000, and the ratio of fluorescence intensity was more than 2 between any two geneprofiles/when the ratio is more than 2,We think the expression is up ,otherwise theexpression is down.Results:1. The reliability of the chip technique: There were 288 cDNA on the chips in comparison with negative rice genome sequence (2 points). The low expression of rice genome by hybridization proved the reliability of data.2. Results of the filtration with gene chip:A. Comparison with esophagcal carcinoma tissue and normal esophagealepitheliumThere were 6 genes expressed up in esophageal carcinoma including: BCL-l, BTK, JNK3A2, CST3, JAW1, Human MHC Class f HLA-A Cell Surface Antigen.There were 8 genes expressed down in esophagea! carcinoma including: ST13, KIAA0161, SOCS-I, D123, PROS-27,1RS1, DRR1, ACTR1A.B. Comparison with pericancerous tissue and normal esophageal epithelium. There were 8 genes expressed up in peri cancerous tissue including: BCL-1, TIAM1, JNK3AI, BTK, IGF1, K-ras, Homo sapiens mRNA for leptin receptor gene-related protein, Human mRNA for hematopoetic proteoglycan core protein. There were 8 genes expressed down in pericancerous tissue including: ST13, PROS-27, IRS1, mucSac, DP-79, APEX, SMAD3, Human colorecta! mutant Cancer protein mRNA.C. Comparison with esophageal carcinoma tissue and pericancerous tissue.There were 10 genes expressed up in esophageal carcinoma including: CD44, int-2,K-ras, R1Z, MRP3, bcl-1, BTK, C-fos, MCAD, Homo Sapiens mRNA for leptinreceptor gene-related protein.There were 5 genes expressed down in esophageal carcinoma including: MCM7,CNNB1, BRF1, TFAP2C, Homo Sapiens mRNA for...