The Screening Of Genes Related To Inhibition Metastasis Of Pancreatic Cancer Under Hypoxia

[Objective] Bioinformatics analyses were used to screen the prognosis-related genes including COL7A1,HIST1H2 BD,MET and SDC1.They are biomarkers with different expression in pancreatic cancer cells before and after hypoxia treatment.Some experiments were used to verify the expression level of these genes in human pancreatic cancer cell(l3.6pl cell)in hypoxia treatment group and control group,and to study the effects of hypoxia treatment on the migration and invasion of pancreatic cancer cells.We aim to explore the role of these genes as important diagnostic and potential therapeutic biomarkers for early development and metastasis of pancreatic cancer.[Methods]1.Bioinformatics analyses were used to analyze differential genes in pancreatic cancer cells before and after hypoxia treatment and to screen out genes related to prognosis of pancreatic cancer.2.Real-time fluorescence quantitative PCR(RT-qPCR)was used to verify the expression leve of COL7A1,HIST1H2 BD,MET and SDC1 genes in human immortalized non-malignant pancreatic cells(HPDE cell line)and human pancreatic ductal carcinoma cells(SW1990 cell line),as well as in human pancreatic cancer cell with high metastatic potential(l3.6pl cell line)before and after hypoxia treatment.3.Scratch test,Transwell migration assay and Transwell invasion assay were used to observe the effect of hypoxia treatment on the migration and invasive ability of human pancreatic cancer cell(l3.6pl cell).[Results]1.There were 298 differentially expressed genes between HPDE cell compared with 20 kinds of pancreatic ductal carcinoma cells and l3.6pl cell under normal compared with hypoxic conditions(P<0.05,F>2).2.These 298 genes were analyzed with GO functional enrichment and KEGG pathway.GO included biological process(BP),cellular component(CC)and molecular function(MF)(P<0.05).3.COL7A1,HIST1H2 BD,MET and SDC1 genes were screened by using twelve algorithms in Cytoscape Hubba,genes’ expression levels of Oncomine database and Kaplan-Meier survival analysis.Compared with normal pancreas tissues,COL7A1,HIST1H2 BD,MET and SDC1 genes are highly expressed in pancreatic cancer.And the expression level of COL7A1,HIST1H2 BD,MET and SDC1 genes are negetively correlated with overall survival(OS)of pancreatic cancer patients,COL7A1 MET and SDC1 genes are negetively correlated with disease-free survival(RFS)(P<0.05).4.The results of PCR(RT-qPCR)showed that the expression levels of COL7A1 and HIST1H2 BD genes in HPDE cells were higher than those in SW1990 cells(P<0.05),which was not consistent with the result from bioinformatics predictions.The results of PCR also showed that the expression levels of MET and SDC1 genes in HPDE cells were lower than those SW1990 cells(P<0.05),which was consistent with the result from bioinformatics predictions.And the expression levels of COL7A1,HIST1H2 BD,MET and SDC1 genes in l3.6pl cell were lower in the hypoxia treatment group than those in the control group(P<0.05),which was consistent with the result from bioinformatics predictions.5.The results of scratch test showed that the wound healing ability in l3.6pl cells in the hypoxia treatment group was lower than that in thecontrol group(P<0.05).Transwell migration experiment results showed that the migration ability in l3.6pl cells in the hypoxia treatment group was lower than that in the control group(P<0.05).Transwell invasion experiment results showed that the invasion ability in l3.6pl cells in the hypoxia treatment group was lower than that in the control group(P<0.05).[Conclusion]1.This research initially screened out MET and SDC1 as target genes related to pancreatic cancer metastasis.2.Hypoxia inhibits metastatic ability of the pancreatic cancer cell.The mechanism may be that hypoxia inhibits the expressions of MET and SDC1 and affects the metastasis way of pancreatic cancer metastasis-related pathways in the hypoxic microenvironment.