High Expression Of Versican In Choroidal Melanoma And Its Mechanism For Promoting Cell Migration And Invasion

Objective: Choroidal melanoma is the most common primary intraocular tumor in adults,with a high degree of malignancy.50% of patients develop metastasis after local treatment,and the one-year survival rate of patients after metastasis is only 15%.Currently,there is no standard treatment for metastatic choroidal melanoma in clinical practice,so it is of great significance to further explore the mechanism of choroidal melanoma metastasis to provide more effective treatment.Versican,encoded by the gene named VCAN,has been widely studied in inflammatory diseases in the early stage.It can participate in extracellular matrix remodeling and enhance inflammatory factor infiltration.In recent years,studies have reported that the expression level of versican in tumor cells is abnormally increased,while it has not been reported in choroidal melanoma.In this study,we will explore the role of versican in choroidal melanoma and its transcriptional regulation mechanism,improve the mechanism of the occurrence and development of choroidal melanoma metastasis,and provide a theoretical basis for finding more effective therapeutic targets.Methods: 1.The expression of VCAN mRNA in patients with uveal melanoma and its influence on prognosis were analyzed by GEPIA and UALCAN databases.2.The expression of versican in choroidal melanoma tissues and non-cancerous choroidal tissues was detected by immunohistochemistry and Western Blot.The expression of VCAN was knocked down by si RNA,and the expression of VCAN mRNA and versican was detected by qRT-PCR and Western Blot,respectively.Wound healing assay,transwell migration and invasion assay were used to detect migration and invasion ability changes.The changes in epithelial-mesenchymal transition(EMT)markers were detected by Western Blot.3.The possibility of FOXA1 transcription regulating VCAN was predicted by bioinformatics websites.The expression of FOXA1 was knocked down by si RNA,and the expression of FOXA1 mRNA and protein was detected by qRT-PCR and Western Blot,respectively.The expression of VCAN mRNA and versican was detected by qRT-PCR and Western Blot,respectively.Wound healing assay,transwell migration and invasion assay were used to detect migration and invasion ability changes.4.The binding sites of FOXA1 and VCAN promoter regions were predicted by bioinformatic websites.Chromatin immunoprecipitation(Ch IP)was used to prove the binding of FOXA1 to the VCAN promoter region.Results: 1.In the public database,VCAN mRNA expression was higher in uveal melanoma patients with high-grade tumors,malignant progression,and shorter survival time.The expression of versican was significantly up-regulated in choroidal melanoma tissues.2.Knockdown of versican resulted in decreased migration and invasion ability of choroidal melanoma cells and decreased expression of EMT mesenchymal markers.3.VCAN mRNA and versican protein expression were significantly down-regulated after FOXA1 knockdown in choroidal melanoma cells.The migration and invasion ability decreased after FOXA1 expression decreased.4.Chromatin immunoprecipitation showed that FOXA1 could bind to the VCAN promoter region.Conclusions: 1.The expression of versican was increased in choroidal melanoma tissues,and was closely related to the disease progression,malignant degree,and poor prognosis of patients.2.Versican promoted the migration and invasion of choroidal melanoma cells and the EMT process.3.The expression of FOXA1 was positively correlated with VCAN and promoted the migration and invasion of choroidal melanoma cells.4.FOXA1 played a transcriptional regulatory role by directly binding to the VCAN promoter.