Molecular Mechanism Of USP10 Regulation Of EIF4G1 Function In Non-Small Cell Lung Cancer

Objectives: Lung tumours are a global health problem and non-small cell lung cancer is the most important category of lung tumours for which there is still a lack of effective early detection indicators and therapeutic targets.Eukaryotic translation initiation factor 4 gamma 1(EIF4G1)is a key component in initiating protein synthesis,and Ubiquitin-specific Protease 10(USP10)is a member of the deubiquitinating enzyme family.They are highly expressed in all types of tumours and have potential value as early diagnostic indicators and therapeutic targets for non-small cell lung cancer.This study explored the regulatory relationship between EIF4G1 and USP10,and also further investigated the role of USP10 in further promoting tumour chemotaxis towards immune cells by affecting EIF4G1.It provides new research ideas for non-small cell lung cancer.Methods: RNA-seq was used to screen for possible molecules downstream of EIF4G1;the expression of various inflammatory factors was examined in lung cancer samples from patients;the intermediate molecule TNFRSF10 A,which regulates CXCL8 by EIF4G1,was identified by immunofluorescence,immunoblotting and fluorescence quantitative PCR;different concentrations of cytokines were added to the lung cancer cell culture medium using enzyme-linked immunosorbent assay.The concentration of CXCL8 in the supernatant of the cell culture medium was measured by enzyme-linked immunosorbent assay(ELISA);the MAPK pathway between TNFRSF10 A and CXCL8 was identified in the literature and verified by immunoblotting;the effect of the tumour on immune cell chemotaxis was examined in a Transwell co-culture system.Results: Previous studies found that EIF4G1 was significantly highly expressed in lung tumors compared to paraneoplastic tissues;USP10binds and regulates EIF4G1 in non-small cell lung cancer cells;transcriptome sequencing screened for EIF4G1 downstream related molecules and chemokine CXCL8 was significantly associated with EIF4G1;applied fluorescence quantitative PCR and ELISA to determine in H1299 cells CXCL8 was highly expressed in H1299 cells at m RNA and protein levels;TNFRSF10A was highly expressed in H1299 stable knockdown EIF4G1 cell line and H1299 stable knockdown USP10 cell line as determined by immunofluorescence,protein immunoblotting and fluorescent quantitative PCR;TNFSF10 was added to H1299 stable knockdown EIF4G1 cell line and H1299 stable knockdown USP10 cell line at different concentrations.The expression of CXCL8 was significantly altered by adding different concentrations of TNFSF10 to the H1299 stable knockdown EIF4G1 cell line and H1299 stable knockdown USP10 cell line;EIF4G1 enhanced the immune cell chemotaxis of H1299 cells by affecting the secretion of CXCL8 as demonstrated by Transwell.Conclusions: 1.USP10 binds to EIF4G1 and the expression level of USP10 is positively correlated with that of EIF4G1 in H1299 cell line.2.EIF4G1 activates MAPK signaling pathway through regulating the expression of TNFRSF10 A to control the secretion of CXCL8,which further affects the chemotaxis of tumor cells to immune cells.