The Regulation Of Wnt/?-catenin Signaling Pathway On Human Tongue Papilla Development And CAL 27 Cell Of Tongue Cancer

The tongue of a mammal has a taste function in acid,sweetness,bitterness,saltiness and freshness,mainly due to the presence of taste papillaes on the surface of the tongue.It has been found that Wnt/?-catenin plays an important role in the early development of mouse tongue papillae,but it is rarely verified during the early development of human tongue papillae.In the first part of the study,the morphological characteristics of human embryonic tongue papillaes were detected,especially on fungiform and circumvallate.The morphology and development process of human tongue papillaes are familiar with those in mouse,but there are some differences in quantity and arrangement.RNA in situ hybridization was used to detect the expression pattern of key genes of Wnt/?-catenin signaling pathway in human tongue embryos.The results showed that Wnt/?-catenin signaling pathway had high activity in early development of human tongue papillae,which always keep changing dynamically.Furthermore,human tongue embryos in different periods were cultured with CHIR-99021 to increase the Wnt/?-catenin signal activity.The results revealed that the number of fungiform on the surface of human tongue was significantly increased in the experimental group;the expression of AXIN2,LEF1,CCND1 and the nuclear activated ?-catenin in the Wnt/?-catenin signaling pathway were significantly up-regulated.Finally,immunofluorescence technique was utilized to detect the progenitor cell marker protein Ki67 and the nerve fibers marker protein PGP9.5.The results showed that the increased of bacterial papillae of human tongue,which was activated by of Wnt/?-catenin signaling,was unrelated to cell proliferation,but increased the amount of taste nerve fiber.Our results demonstrated that CHIR-99021 can increase the activity of Wnt/?-catenin signaling in human tongue embryos and increase the number of tongue papillaes.In addition,it has been reported that the ?-catenin is abnormally expressed in tongue cancer tissues.Adult tongue cancer tissues and adjacent normal tissues were collected in the second part of the study.The expression of ?-catenin in tongue cancer tissues was confirmed by immunohistochemistry and Western Blot.Secondly,tongue cancer cell line CAL 27 was used to investigate the effects of FH535,a Wnt/?-catenin signaling inhibitor on tongue cancer cells.The results showed that cell proliferation,migration and clonality were inhibited in CAL 27 cells which were treated with FH535.But the apotheosis rate was significantly increased.The expression of E-cadherin,an epithelial marker of epithelial-mesenchymal transition(EMT)was up-regulated,while N-cadherin,a cytoplasmic marker was down-regulated.In summary,Wnt/?-catenin signaling pathway plays an important role in the early development of human tongue.Its abnormal expression is related to the formation of tongue cancer,and its signal activity is important to inhibit the development of tongue cancer cells.