Expression Of C-Fos In Oral Squamous Cell Carcinoma And Its Effect On Growth And Invasion

Object: To identify the expression of c-Fos in OSCC tissues and cells and its association with the clinical characteristics and prognosis,then search its functions in proliferation,migration and invasion in OSCC cells.We predict miR-155-5p may target regulating c-Fos through the analysis of the bioinformatics,verify whether the miR-155-5p could target to regulate c-Fos in OSCC.To provide some experimental and theoretical basis for finding the treatment target of OSCC and improving the treatment and prognosis of patients with OSCC.Methods: Bioinformatics was used to analyze the expression of c-Fos in OSCC and its correlation with clinical parameters,and predict the miRNA that may target and regulate c-Fos.Immunohistochemistry(IHC)was utilized to detect the expression of c-Fos in 94 OSCC tissues and 30 paracancerous tissues,then explore the connection between c-Fos and the clinicopathological characteristics of OSCC patients and the prognostic value.The relative content of c-Fos in OSCC cells was examined by the real-time quantitative PCR(qRTPCR)and Western Blot,and then the cell experiment in vitro was carried out: transfected OSCC cells(SCC15,CAL27)with c-Fos overexpression plasmid(GV657-c-Fos),qRT-PCR and Western Blot were utilized to estimate the transfection efficiency.After successful transfection,we estimated the growth and invasion of c-Fos in OSCC cells by Colony formation assay,CCK-8 assays,Wound healing assay and Transwell assays.In addition,based on bioinformatics prediction,after transfection of OSCC cells(SCC15,CAL27)with miR-155 inhibitor or mimic,qRT-PCR assessed the transfection efficiency,then qRT-PCR and Western Blot were used to test the regulation of miR-155-5p on c-Fos expression,then the Dual-luciferase reporter gene assays was utilized to verify whether miR-155-5p regulates the transcription activity of c-Fos in OSCC.Results:1.GEO database showed that c-Fos was highly expressed in OSCC(P<0.01),and bioinformatics prediction analysis miR-155-5p may target and regulate c-Fos in OSCC.2.IHC showed that c-Fos was highly expressed in OSCC tissues(P<0.01).c-Fos expression in OSCC tissues was considerably related to lymph node metastasis(P<0.01)and differentiation(P<0.01).Nevertheless,there was no significant connection between the expression of c-Fos and the clinical stage(P>0.05),TNM stage(P>0.05),age(P>0.05)and sex(P>0.05).In addition,c-Fos expression was negatively connected with overall survival(OS)(P<0.01)and the disease-free survival(DFS)(P<0.01).3.The outcome of qRT-PCR(P<0.01)and Western Blot(P<0.05)exhibited that c-Fos was overexpressed in OSCC cells.After overexpression of c-Fos,The Colony formation assay(P<0.01)and the CCK8 assay(P<0.05)confirmed that the growth and proliferation of SCC15 and CAL27 cells increased;the Wound healing assay(P<0.01)and Transwell assays(P<0.05)indicated that the migration and invasion ability of SCC15 and CAL27 cells was enhanced.4.The qRT-PCR(P<0.05),Western blot(P<0.01)and the Dual-luciferase reporter gene assay(P<0.01)revealed miR-155-5p regulated the transcription activity of c-Fos by directly targeting 3’-UTR of c-Fos in OSCC,thus positively target regulating c-Fos.Conclusion: c-Fos expression is upregulated in OSCC tissues and cells.c-Fos is connected with lymph node metastasis,differentiation and poor prognosis of OSCC patients,c-Fos promotes the proliferation,migration and invasion in OSCC cells.This study preliminarily verified that miR-155-5p is positively target regulating c-Fos in OSCC.c-Fos may be a potential prognostic marker and a new therapeutic target for OSCC.