Abstract
BackgroundTo screen and analyze differentially expressed genes in pancreatic carcinoma tissues taken from Mongolian and Han patients by Affymetrix Genechip. Methods: Pancreatic ductal cell carcinoma tissues were collected from the Mongolian and Han patients undergoing resection in the Second Affiliated Hospital of Nanchang University from March 2015 to May 2018 and the total RNA was extracted. Differentially expressed genes were selected from the total RNA qualified by Nanodrop 2000 and Agilent 2100 using Affymetrix and a cartogram was drawn; The gene ontology (GO) analysis and Pathway analysis were used for the collection and analysis of biological information of these differentially expressed genes. Finally, some differentially expressed genes were verified by real-time PCR.ResultsThrough the microarray analysis of gene expression, 970 differentially expressed genes were detected by comparing pancreatic cancer tissue samples between Mongolian and Han patients. A total of 257 genes were significantly up-regulated in pancreatic cancer tissue samples in Mongolian patients; while a total of 713 genes were down-regulated. In the Gene Ontology database, 815 differentially expressed genes were identified with clear GO classification, and CPB1 gene showed the highest increase in expression level (multiple difference: 31.76). The pathway analysis detected 28 signaling pathways that included these differentially expressed genes, involving a total of 178 genes. Among these pathways, the enrichment of differentially expressed genes in the FAK signaling pathway was the strongest and COL11A1 gene showed the highest multiple difference (multiple difference: 5.02). The expression of differentially expressed genes CPB1, COL11A1、ITGA4、BIRC3、PAK4、CPA1、CLPS、PIK3CG and HLA-DPA1 determined by real-time PCR were consistent with the results of gene microarray analysis.ConclusionsThe results of microarray analysis of gene expression profiles showed that there are a large number of differentially expressed genes in pancreatic cancer tissue samples comparing Mongolian and Han population. These genes are closely related to the cell proliferation, differentiation, invasion, metastasis and multi-drug resistance in pancreatic cancer. They are also involved in the regulation of multiple important signaling pathways in organisms.
Highlights
Pancreatic cancer has the highest mortality among all the digestive system malignancies
Reagents and instruments The required reagents Agilent Ribonucleic Acid (RNA) 6000 Nano Kit, GeneChip 3’IVT Express Kit, GeneChip Hybridization Wash and Stain Kit, Trizol Kit, and QIAGEN RNeasy Total RNA Isolation kit were provided by Gekkai Gene Technology Co., Ltd
Sample information A total of 96 fresh tumor tissue specimens from patients with pancreatic malignancies surgically resected in the Second Affiliated Hospital of Nanchang University and the First Affiliated Hospital of Nanchang University from March 2015 to May 2018 were collected. 66 of them were initially selected according to the following inclusion and exclusion criteria and these specimens were used for follow-up gene expression microarray assays
Summary
Pancreatic cancer has the highest mortality among all the digestive system malignancies. By analyzing genes involved in the generation and maintaining of malignant biological characteristics of pancreatic cancer, this method [2] can elucidate the molecular mechanism of pancreatic cancer. Affymetrix Gene Expression Profiling microarray was used to screen differentially expressed genes in pancreatic cancer samples of Mongolian and Han patients for further gene function analysis, in order to provide important reference data and experimental evidence for the elucidation of the molecular mechanism of pancreatic cancer development. To screen and analyze differentially expressed genes in pancreatic carcinoma tissues taken from Mongolian and Han patients by Affymetrix Genechip. Methods: Pancreatic ductal cell carcinoma tissues were collected from the Mongolian and Han patients undergoing resection in the Second Affiliated Hospital of Nanchang University from March 2015 to May 2018 and the total RNA was extracted. Some differentially expressed genes were verified by real-time PCR
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