FOXK1通过上调ROCK1表达促进食管鳞状细胞癌的恶性生物学行为OA

Objective To investigate the expression,biological function,and underlying molecular mechanisms of FOXK1 in esophageal squamous cell carcinoma(ESCC).Methods ESCC cell lines KYSE-150,KYSE-170,and TE-1 were routinely cultured.Nucleic acids and plasmids were transfected into cells using transfection reagents.Data-base analysis combined with qRT-PCR was applied to determine FOXK1 expression in ESCC tissues and cells.Bioin-formatics analysis was used to predict downstream targets,with ROCK1 identified as a candidate.ROCK1 expression was examined by qRT-PCR,and a dual-luciferase reporter assay was proformed to confirm FOXK1-mediated transcrip-tional regulation of the ROCK1 promoter.Colony formation,wound-healing,and Transwell assays were conducted to evaluate cell proliferation,migration,and invasion.qRT-PCR and Western blotting were used to examine the expres-sion of epithelial-mesenchymal transition(EMT)-related markers,while immunofluorescence was applied to assess F-actin polymerization.Results FOXK1 expression was significantly upregulated in ESCC tissues and cell lines.In KYSE-150,KYSE-170,and TE-1 cells,FOXK1 expression was approximately 40.0-,17.9-,and 24.8-fold higher than that in normal esophageal epithelial cells,respectively(P<0.05).ROCK1 expression positively correlated with FOXK1 levels(P<0.05),and was also elevated in ESCC cells.FOXK1 overexpression increased,whereas FOXK1 knockdown decreased ROCK1 expression.Dual-luciferase reporter assays confirmed that FOXK1 directly bound to and activated the ROCK1 promoter.Functionally,ROCK1 overexpression enhanced ESCC cell proliferation,migration,and invasion,while ROCK1 knockdown inhibited these phenotypes.Co-manipulation of FOXK1 and ROCK1 partially reversed the pro-tumor effects of FOXK1 overexpression.Mechanistically,ROCK1 knockdown down-regulated the mRNA and protein expression of EMT markers(CDH2,vimentin,and ZEB1).FOXK1 overexpression promoted F-actin polymerization,whereas ROCK1 knockdown suppressed it;co-manipulation partially counteracted FOXK1-induced F-actin polymerization.Conclusion FOXK1 promotes ESCC cell migration and invasion by tran-scriptionally upregulating ROCK1,indicating that FOXK1 may serve as a potential diagnostic and therapeutic target in ESCC.