Bioinformatics Analysis Of Whole Exome Sequencing Data Of Adenocarcinoma Of Esophagogastric Junction And Effects Of Involucrin On The Biological Function Of Tumor Cells

Objective:To detect the gene mutations in patients with adenocarcinoma of the esophagogastric junction(AEG)and analyze the effect of Involucrin(IVL)mutation on the biological function of tumor cells.Methods:1.Whole genome sequencing and IVL mutation analysis of AEG patients in Shanxi. Collect the cancer tissues and matched adjacent normal tissues of 22 patients with AEG who were resected by general surgery in Shanxi Cancer Hospital.Use Next Generation Sequencing(NGS)technology to sequence their entire genomes,analyze the IVL mutations and screen the sites.The IVL mutations of esophageal cancer in the TCGA database were analyzed for function enrichment.2.The effect of overexpression of IVL in KYSE150 on cell function. Human IVL overexpressed plasmid was transfected into KYSE150.Western Blot and q RT-PCR were undertaken to study the overexpression efficiency.MTT and Wound Healing assays were used to evaluate the effect of IVL on the proliferation and migration abilities of KYSE150 cells.3.The binding ability of mutated oligopeptides to transglutaminase 1(TGase1). The binding activity of IVL oligopeptides with different mutation sites strains to transglutaminase 1(TGase1)was analyzed using in vitro activity experiments.Results:Total exon sequencing of cancer and paracancerous tissues from 22 patients with AEG in Shanxi revealed that the top 10 genes with mutation rates were TP53,TTN,MUC16,NALCN,IVL,LAMA5,NEB,OBSCN,PCLO,and ROBO1.There were 5 mutations in 4AEG tumor samples,all of which were missense mutations.Sequencing showed that the mutation rate of IVL gene in AEG patients in Shanxi region was as high as 18%.GO annotation indicated that these mutations were related with the epithelial differentiation and activity of cell differentiation regulation.KEGG analysis demonstrated that IVL mutations were associated with c AMP signaling pathway,etc.IVL overexpression could inhibit the proliferation and migration of esophageal cancer cells.Besides,Q439 H mutant IVL oligopeptides showed lower binding ability with TGase1 than wild-type oligopeptides.Conclusion:Mutations occurred at the seventh amino acid in IVL repeat motif could weaken IVL binding with TGase1.The mutations may affect tumor-related biological processes and signaling pathways.