Bioinformatics Analysis Of SPAG4 Pan-Cancer And Its Expression Validation In Osteosarcoma

Purpose:Osteosarcoma(OS),as the most common primary bone malignancy,is very harmful to patients,and the search for new therapeutic targets in order to improve the prognosis of patients remains one of the current research hotspots.The expression of sperm associated antigen 4(SPAG4)as a tumor marker in osteosarcoma has not been reported in the literature.In this study,we intend to conduct a preliminary investigation on the expression of SPAG4 in osteosarcoma and its mechanism through bioinformatics and related experiments.Methods:(1)The expression levels of SPAG4 in 33 tumors and its relationship with tumor prognosis were investigated by bioinformatics analysis of TCGA,GEO and GTEx data.(2)The analysis of TARGET database was used to explore whether SPAG4 has differential mRNA expression in osteosarcoma and normal tissues,and to assess the differential prognosis of osteosarcomatized patients in the high and low SPAG4 expression groups.The protein expression levels of SPAG4 in osteosarcoma tissues and cell lines were detected by immunohistochemistry,protein immunoblotting technique and immunofluorescence.(3)To analyze the interacting proteins of SPAG4 by String web tool,and to perform KEGG pathway analysis and GO function analysis of SPAG4 by DAVID web tool.(4)Preliminary exploration of the mechanism of SPAG4 in osteosarcoma using immunofluorescence,protein immunoblotting and immunoprecipitation techniques.Resullts:(1)SPAG4 is highly expressed in a variety of human tumors,and high SPAG4 expression is associated with poor clinical prognosis of patients.(2)Biochemical analysis of osteosarcoma-related data from TARGET and GTEX databases revealed that SPAG expression was upregulated in osteosarcoma.(3)We verified that SPAG4 was significantly overexpressed in osteosarcoma by immunohistochemistry and other experiments.(4)Enrichment analysis of SPAG4-interacting proteins and SPAG4-related genes revealed the possible interaction of SPAG4 with CCNB1 in the P53 signaling pathway.(5)The possible interaction of SPAG4 with CCNB1 is involved in the development of osteosarcoma.Conclusions:(1)SPAG4 is highly expressed in a variety of tumors and is a tumor prognostic marker that can be used as a marker to assess prognosis.(2)SPAG4 is significantly highly expressed in osteosarcoma(3)SPAG4 interacts with CCNB1 and has the potential to play a role in the development of osteosarcoma.