A Primary Study On The Role Of Wnt5a In Skin Epithelium

At the surface of our body, the skin epithelium provides a protectivebarrier by undergoing continual self-renewal. The epidermis undergoeshomeostatic regulation as basal cells periodically execute their programmeof terminal differentiation and move outwards in a columnar fashion. Hairfollicle is an important and complex appendage in the skin. The hair follicleunit consist of the epithelium cell and mesenchyme cell. It keeps on growthand self-renewal by periodic hair cycle. There are multiple signallingpathways which are are essential for the epidermis and hair follicle. As atypical noncanonical Wnt, Wnt5a plays an important role in skin woundhealing. Recently, Wnt5a can inhibit hair shaft growthin vitro. However,the role of Wnt5a to skin epithelium are not well understand.In our study, the research on the effect of Wnt5a on the proliferation ofmouse epidermal cell was firstly detected by cell counting and BrdUincorporation analysis. Secondly, we analyzed changes in non-canonicalWnt pathway signaling molecules by RT-PCR. Finally, the key moleculeswhich is significantly different were screed and its expression in theepithelium of the skin was detected. This study results are as follows:1. AdWnt5a-GFP, AdGFP, AdWnt10b were amplified in HEK293cells. 2. Used “JB6” as a cell model in vitro, and infected it withAd-Wnt5a-GFP or Ad-GFP at a pre-determined optimal titer.3. The effect of Wnt5a on JB6proliferation was evaluated by cellcount assay and BrdU incorporation.4. The effect of Wnt5a on JB6differentiation was evaluated byRT-PCR.5. The ability of Ad-Wnt5a-GFP to inhibit Wnt/β-catenin signalingwas evaluated by RT-PCR and Western blot.6. The ability of Ad-Wnt5a-GFP to activate noncanonical Wntsignaling was evaluated by RT-PCR and Western blot.7. The expression profile of JNK was detected by IHC in the skinepidermis in postal mouse. JNK was expressed in the epidermis.8. The expression profile of JNK and Wnt5a was detected by IHC inthe skin epidermis in human skin. Wnt5a was expressed in the basal cellsand superbasal cells in the epidermis. And JNK was in the superbasal cells.9. The expression profile of JNK was detected by IHC in hair cycle inpostal mouse. JNK is dynamic factor in hair cycle.10. The expression profile of JNK was detected by RT-PCR and WBin hair cycle. The results of IHC，RT-PCR and WB are consistent.In conclusion, the research explored the role of Wnt5a in the skinepithelia.Wnt5a inhibites the proliferation and differentiation of JB6cell,and activates noncanonical Wnt signaling to antagonize canonical Wnt signaling. In addition, as downstream signaling molecules, the expressionof JNK is similar with Wnt5a. Our study provides a new theory for thefurther study of the mechanism of action of Wnt5a in the skin.