SPATS2 promotes LSCC process by activating the NOTCH signaling pathway
Objective:This study aims to investigate the molecular mechanisms through which spermatogenesis associated ser-ine rich 2(SPATS2)influences larynx squamous cell carcinoma(LSCC)by affecting proliferation,migration,invasion,and epithelial-mesenchymal transition(EMT).The findings will provide a theoretical basis for identifying potential targets for screen-ing,diagnosis,and treatment LSCC.Methods:Clinical and RNA-seq data for LSCC were obtained from the TCGA database and processed accordingly.SPATS2 mRNA expression data were extracted and analyzed using R 4.3.1 software.Gene ontology(GO)analysis and gene set enrichment analysis(GSEA)were conducted using the gene set obtained from single-gene differential analysis performed with R version 4.3.1.Proliferation,migration,and invasion abilities of cells were assessed through CCK8,col-ony formation,wound healing and Transwell assays.Western blotting was employed to assess protein expression,while qRT-PCR was used to measure mRNA levels of genes.Quantitative and statistical analyses were performed using ImageJ,Graph-Pad Prism 8,and IBM SPSS Statistics 25.Results:Bioinformatics analysis revealed significantly higher levels of SPATS2 mRNA in LSCC tissues compared to adjacent normal tissues(P<0.001),indicating an adverse impact on patient survival(P=0.021).Overexpression or knockdown of SPATS2 resulted in altered cell proliferation,migration,invasion,as well as changes in the expression levels of Vimentin,N-Cadherin,NOTCH1 and cleaved-NOTCH1,and downstream target genes of the NOTCH signaling pathway.Inducing overexpression of SPATS2 and inhibiting the NOTCH signaling pathway simultaneously could re-verse the enhanced cell proliferation,migration,invasion,and EMT caused by SPATS2 overexpression.Conclusion:SPATS2 exerts influence on the malignant characteristics of LSCC cells,including proliferation,migration,invasion,and EMT,by modu-lating the NOTCH signaling pathway,thus contributing to the progression of LSCC.