Analysis Of The Correlation Between High ALYREF Expression And Poor Prognosis Of Esophageal Cancer

Objective: ALYREF regulates the nuclear export of m RNA as part of the constituent TREX complex.Recent studies have confirmed the involvement of ALYREF in the development and progression of several human cancers,and its role in esophageal cancer remains unknown.The aim of this study is to analyse the effect of ALYREF on the role of esophageal cancer and to provide new targets for the treatment of esophageal cancer.Methods: This project will be carried out in two parts as follows:1.Conducting bioinformatics analysis.Firstly,the online database TIMER was used to analyse the differential expression of ALYREF in pan-cancer,download and collate transcriptome data from esophageal cancer and compare the differential expression of ALYREF in esophageal cancer with adjacent normal tissues.The differential genes were screened to draw volcano maps and gene correlation heat maps.The correlation between ALYREF and clinicopathological features,its impact on patient prognosis were then analysed,and univariate and multifactorial Cox regression analyses were used to determine whether ALYREF was an independent prognostic factor and its value as a prognostic indicator was assessed.Subsequently,GO(Gene Ontology),KEGG(Kyoto Encyclopedia of Genes and Genomes)and GSEA(Gene Set Enrichment Analysis)pathways were analysed to explore The main pathways of action of ALYREF were explored.The correlation between ALYREF expression and the level of immune cell infiltration in human 24 and the correlation with immune regulatory genes were then explored.All raw data were downloaded from TCGA(The Cancer Genome Atlas)and analysed and visualised using version R(3.6.3).2.In vitro cell function assays and immunohistochemistry were performed.Two esophageal cancer cell lines,KYSE150 and ECA109,were selected and transfected with lentivirus to knock down the expression level of the ALYREF gene.Cell cycle assays,CCK8 assays and scratch assays were performed to observe whether cell cycle alteration occurred after gene knockdown and affected the proliferation and migration ability of the cells.After patients’ informed consent,post-operative tissue samples from clinical esophageal cancer patients were collected and immunohistochemical assays were performed to observe the staining of ALYREF in oesophageal cancer tissues and paracancerous tissues.Results:1.ALYREF was significantly highly expressed in esophageal cancer and 18 other human cancers,where the overall survival(OS),disease-specific survival(DSS)and progression-free survival(PFI).Cox regression results showed that N stage,M stage,treatment outcome,ethnicity,tumour residual and ALYREF were all independent prognostic factors for patients(p < 0.05).Enrichment analysis showed that differential genes were significantly enriched in pathways such as cell cycle,unwinding helicase activity and DNA replication.Moreover,ALYREF was significantly and positively correlated with the expression of various cell cycle-regulated genes in vivo,suggesting that ALYREF may promote the progression of oesophageal cancer by affecting the cell cycle.In addition,the high expression of ALYREF was also negatively correlated with the infiltration level of various immune cells,chemokines and immune checkpoints in vivo.2.In vitro experiments to knock down the expression of ALYREF showed that the knockdown of ALYREF resulted in S-phase arrest and inhibition of proliferation and migration of KYSE150 and ECA109 cells.Immunohistochemical results confirmed that ALYREF was more highly expressed in the patient’s esophageal cancer tissues than in the adjacent normal tissues.Conclusion: In this study,bioinformatics analysis combined with in vitro functional assays revealed that ALYREF expression was significantly increased in esophageal cancer tissues and correlated with poor prognosis of patients.It is suggested that ALYREF plays an important role in esophageal cancer and may be a prognostic biomarker for esophageal cancer.