Screening Differential Expression Genes With Different Syndromes In Human Esophageal Squamous Cell Carcinoma

Objective:To study the related gene expression in esophageal carcinoma of different stages(cancerous tissue, paraneoplastic tissue, normal tissue) by cDNA microarray technique,screen differentially expressed genes of esophageal cancer tissue in different syndromes,preliminarily reveal the correlations of gene expression in different syndromes ofesophageal cancer, and analyze their possible existing molecular signal transmissionmechanism.Methods:①Tissue specimen database and clinical information database of esophagealcancer were established;21cases of cancerous tissue, paraneoplastic tissue, normalmucosa tissue of esophageal cancer from rigidly matched surgical tissue specimens werescreened; Then successively compared the gene expression spectrum in proper sequence of21esophageal cancer cases of cancerous tissue, paraneoplastic tissue, normal mucosatissue, and of different syndromes with stagnation of sputum and qi syndrome (P),stagnation of stasis blood syndrome (B), insufficiency of body fluid and agglomeration ofheat syndrome (F), insufficiency of qi and yang syndrome (Q).②The whole RNA of21cases of cancerous tissue, paraneoplastic tissue, normal mucosa tissue of esophageal cancerwas extracted; agarose gel electrophoresis was made quality control; total RNA waspurified and reversely transcribed into cDNA, and cancerous tissue, paraneoplastic tissue,normal mucosa tissue of esophageal cancer were marked by single notation of Cy3-dUTPso that to manufacture probe, which was hybridized with cDNA microarray. Then scanningfluorescence signal image of gene chip and using SAS software to analyze the microarrayimage to select out differential expression genes.Results:①The following results were analyzed by using SAS software: Cancer tissue andparaneoplastic tissue had2505kinds of differentially expressed genes, Cancer tissue andnormal tissue had2659kinds of differentially expressed genes. In the group P and thegroup B, there were18kinds of differentially expressed genes in cancerous tissue,10inparaneoplastic tissue,6in normal tissue; In the group P and the group F, there were52 kinds of differentially expressed genes in cancerous tissue,8in paraneoplastic tissue,2innormal tissue; In the group P and the group Q, there were52kinds of differentiallyexpressed genes in cancerous tissue,24in paraneoplastic tissue,46in normal tissue; In thegroup Q and the group B, there were12kinds of differentially expressed genes incancerous tissue,5in paraneoplastic tissue,84in normal tissue; In the group Q and thegroup F, there were26kinds of differentially expressed genes in cancerous tissue,6inparaneoplastic tissue,8in normal tissue; In the group B and the group F, there were22kinds of differentially expressed genes in cancerous tissue,8in paraneoplastic tissue15in normal tissue.②The screened differential expression genes in different stages ofesophageal carcinoma were mainly divided into three categories according to GO Category,namely, cellular component,molecular function and biological process. If divided bymolecular function, most of these genes were relevant to catalytic activity, signaltransduction, molecular transporter, activity regulation, protein transport function, cellgrowth and apoptosis.③We found that several major Pathways in different stages ofesophageal carcinoma were involved in Neuroregulin receptor degredation protein-1Controls ErbB3receptor recycling, Inhibition of Matrix Metalloproteinases, SmallLeucine-rich Proteoglycan (SLRP) molecule, Mechanism of Acetaminophen Activity andToxicity, BRCA1-dependent Ub-ligase activity, RB Tumor Suppressor/CheckpointSignaling in response to DNA damage, Role of Ran in mitotic spindle regulation, DNAreplication-Homo sapiens (human); With analyzing the signal pathway of screeneddifferential expression genes in different syndromes of esophageal carcinoma, we foundthat the signal pathway of differential expression genes existing in the group P and thegroup B mainly were TSP-1Induced Apoptosis in Microvascular Endothelial Cell,Drugmetabolism-cytochrome P450-Homo sapiens and Drug metabolism other enzymes Homosapiens (human); The signal pathway of differentially expressed genes existing in thegroup P and the group F mainly related with the pathways of Dendritic cells in regulatingTH1and TH2Development,Sonic Hedgehog (SHH) Receptor Ptc1Regulates cell cycleand ADP-Ribosylation Factor; The signal pathways of differential expression genesexisting in the group P and the group Q mainly were Sonic Hedgehog (SHH) Receptor Ptc1Regulates cell cycle, Leukocyte transendothelial migration-Homo sapiens (human),Antigen Dependent B Cell Activation, Alternative Complement Pathway, SmallLeucine-rich Proteoglycan (SLRP) molecules; The signal pathways of differentialexpression genes existing in the group Q and the group B mainly were Rab GTPases MarkTargets In The Endocytotic Machinery,Drug metabolism-cytochrome P450-Homo sapiens (human), Transcriptional activation of dbpb from mRNA, Classical Complement Pathway,Complement Pathway; The signal pathways of differential expression genes existing in thegroup Q and the group F mainly were Limonene and pinene degradation-Homo sapiens(human),Th1/Th2Differentiation, The Co-Stimulatory Signal During T-cell Activation,Phospholipase C-epsilon pathway; The signal pathways of differential expression genesexisting in the group B and the group F mainly were Bone Remodelling,IL17SignalingPathway, Drug metabolism-cytochrome P450-Homo sapiens (human), Endocytosis-Homosapiens (human).Conclusion:①Gene expression profiles at different stages of esophageal cancer weredifferent. Neuroregulin receptor degredation protein-1Controls ErbB3receptor recyclingPathway、 Inhibition of Matrix Metalloproteinases pathway and Cell cycle signaltransduction pathway for esophageal cancer three major signal transduction pathways②Gene expression profiles in different TCM syndromes of esophageal cancer were different.the function of differentially expressed gene in the mutual stagnation of sputum and qisyndrome mainly was to DNA metabolic process、cell proliferation、DNA replication andactivity regulation; the function of differentially expressed gene in the insufficiency ofbody fluid and agglomeration of heat syndrome mainly was to DNA metabolic process、thephase of mitosis、phosphorus metabolism; the function of differentially expressed gene inthe internal stagnation of stasis blood syndrome mainly was to mitotic cell cycle、nucleardivision、cells division、cell proliferation; the function of differentially expressed gene inthe insufficiency of qi and yang syndrome mainly was to induced cell proliferation andmigration、phosphorylation、protein transport；③We found that several major Pathways indifferent syndromes of esophageal carcinoma,the Cell cycle signal transduction pathwaysall involved in four different syndromes of esophageal cancer, the differentially expressedgene in the mutual stagnation of sputum and qi syndrome mainly was involved in MAPKsignaling pathway; the differentially expressed gene in the insufficiency of body fluid andagglomeration of heat syndrome mainly was involved in participate in the Ca2+signaltransduction and gene transcription; the function of differentially expressed gene in theinternal stagnation of stasis blood syndrome mainly was involved in participate in P53signal transduction and DNA replication signaling pathwayl; the function of differentiallyexpressed gene in the insufficiency of qi and yang syndrome mainly was involved in ErbBsignaling pathway and cell phagocytic pathway...