Study On The Role Of Arecoline Inhibits MiR-203a-5p Through Targeting PTK2 In Promoting BMFs Activation

Background/Purpose: Oral Submucosal Fibrosis(OSF)is a disease with a slow process,which can transform into oral squamous cell carcinoma and mainly occurs in oral buccal mucosa.The incidence was related to betel nut chewing,although the pathogenic mechanism remains unknown.Numerous studies have demonstrated that numerous micro RNAs are related with the development of oral submucosal fibrosis(OSF),a precancerous lesion of the oral cavity.In the early stage,our research group found that the expression of mir-203a-5p in OSF was significantly down regulated through Affymetrix mi RNA chip experiment The expression of mi R-203a-5p is dysregulated in various cancer,however its significance in OSF has not been explored.The purpose of the study is to examine the functional role of mi R-203a-5p and its target in OSF.Methods: The expression levels of mi R-203a-5p in fibrotic buccal mucosal fibroblasts(f BMFs)from OSF tissues and stimulation of BMFs with various concentration of arecoline were examined using nextgeneration sequencing and real-time Polymerase Chain Reaction(PCR)analysis.Mi R-203a-5p mimics were applied to study its functional involvement in proliferation ability of arecoline-treated BMFs.In addition,many myofibroblast characteristics,such as migration,wound healing and collagen gel contraction were examined,and a luciferase reporter experiment was undertaken to demonstrate the link between mi R-203a-5p and its anticipated target PTK2.The expression of α-SMA and PTK2 were assessed in f BMFs from OSF tissues and in the arecoline-induced BMFs to figure out their association between OSF.Meanwhile,we investigated the impact of mi R-203a-5p on PTK2.Results: We detected down-regulation of mi R-203a-5p in f BMFs from OSF and in arecoline-induced BMFs.Arecoline promotes the transdifferentiation of fibroblasts into myofibroblasts.However,transfection of mi R-203a-5p mimics decreased the migratory ability and collagen gel shrinkage of BMFs produced by arecoline.Furthermore,we also demonstrated that the expression of a-SMA and PTK2 was upregulated in f BMFs and in arecoline-induced BMFs,and their expression were decreased after transfection of mi R-203a-5p mimics.The results of the luciferase reporter assay indicated that PTK2 was a direct target of mi R-203a-5p.Conclusion: Arecoline promotes myofibroblast activation,and mi R-203a-5p can inhibit arecoline induced myofibroblast activation.There is a targeted regulatory relationship between mir-203a-5p and PTK2.