The Role And Mechanism Of SPICE1 In Osteosarcoma

Background and Objective:Osteosarcoma is the most common primary malignant bone tumor,which mostly occurs in children and adolescents,with a global incidence of approximately3.4 cases per million a year.Although the techniques and methods of osteosarcoma treatment have been improving,the prognosis of osteosarcoma patients has been relatively stagnant since the advent of modern chemotherapy in the 80 s of the 20 th century.Therefore,it is of great significance to deeply explore the molecular regulatory mechanism of osteosarcoma occurrence and development and actively find new therapeutic targets to improve the prognosis of osteosarcoma patients.In our previous study,we collected three pairs of osteosarcoma and para-tumor tissues for RNA sequencing and intersected the differentially expressed genes with the genes related to osteosarcoma prognosis in the TARGET database,and then screened out SPICE1 as the target gene.SPICE1(Spindle and centriole-associated protein 1)is a centriole and spindle-associated protein whose function is mainly involved in regulating the number and length of centrioles and playing a role in spindle tissue.However,SPICE1 has not been studied in tumors.In this study,we find that SPICE1 was highly expressed in osteosarcoma,and besides,SPICE1 was positively correlated with poor prognosis in patients with osteosarcoma.Therefore,we hypothesized that SPICE1 may play an important role in the progression of osteosarcoma.In order to test this conjecture,this study intends to explore the role and molecular mechanism of SPICE1 in osteosarcoma via in vivo and in vitro experiments.Methods:Firstly,Western Blotting and RT-q PCR experiments were used to detect the expression of SPICE1 protein and m RNA in osteosarcoma and paratumor tissues,osteosarcoma cell lines and human osteoblast h FOB 1.19.Secondly,SPICE1overexpression/interference lentiviral vectors were used to construct overexpression/interference stable cell lines,and the biological function of SPICE1 in osteosarcoma was detected by CCK8,clonal formation,Transwell invasion and migration experiments and subcutaneous tumor experiments.Finally,the IP-MS technology was used to explore the protein interacting with SPICE1,and the molecular mechanism of SPICE1 promoting the progression of osteosarcoma was explored through Co-IP experiment,cellular immunofluorescence experiment,ubiquitination experiment,protein degradation experiment,etc.Results:1.SPICE1 is highly expressed in osteosarcoma tissues and cells and it is positively correlated with poor prognosis in osteosarcoma patients.2.SPICE1 promotes the proliferation,invasion and migration of osteosarcoma cells,and enhances the subcutaneous tumorigenesis ability of osteosarcoma cells in nude mice.3.SPICE1 interacts with FASN,and besides,SPICE1 positively regulates FASN protein levels without affecting its m RNA levels.It is found that SPICE1 stabilizes FASN by inhibiting FASN ubiquitination degradation.4.USP10 was screened out by intersecting the IP-SPICE1,IP-FASN and deubiquitinase datasets.Subsequently,it was confirmed that USP10 interacted with SPICE1 and FASN,and that SPICE1 recruited USP10 to inhibit ubiquitination degradation of FASN.5.Interference with USP10 or FASN inhibits proliferation,invasion,migration and subcutaneous tumorigenesis of osteosarcoma cells promoted by SPICE1.Conclusions:1.SPICE1 is highly expressed in osteosarcoma and it is positively correlated with the poor prognosis of osteosarcoma patients.2.SPICE1 promotes the proliferation,invasion,migration and subcutaneous tumorigenesis of osteosarcoma cells in nude mice.3.SPICE1 recruits USP10 to stabilize FASN and promote osteosarcoma progression.