Effect Of Annexin A2 Protein On Dermal Papilla Cells And Its Mechanism Of Action In Wnt Signaling Pathway

Background: The imbalance of hair follicle self-renewal ability and the abnormality of periodic growth may lead to hair loss,hairy and even tumor.Therefore,it is of great clinical significance to study the regulation signal of hair follicle growth.The Wnt/?-catenin signaling pathway is a central signaling pathway regulating the formation of embryonic hair follicle and the development of adult hair follicle,which plays a very important role in the growth and development of hair follicle,the maintenance and activation of hair follicle stem cells,the color and morphology of hair,and the formation of skin tumors.Dermal papilla cells(DPC)are an important part of hair follicles,and its aggregative growth plays an important role in maintaining and inducing hair follicle regeneration.In our previous study,We have identified 28 proteins that were differentially expressed,including Annexin A2,in aggregative and non-aggregative growth of dermal papilla cells.Aim: To investigate the effect of Annexin A2 on DPC and its mechanism of action in Wnt signaling pathway.Methods: We detected the differences in proliferation and apoptosis between the passage 3 and the passage 10 DPC by flow cytometry and CCK-8.Suppressed the expression of Annexin A2 in passage 3 DPC and up-regulated the expression of Annexin A2 in passage 10 DPC by constructing adenoviral vector of Annexin A2.We used q RT-PCR and Western Blotting to detect the expression level of Annexin A2,apoptotic factor Bcl-2 and proliferative factor PCNA between the passage 3 and the passage 10 DPC,the knock-down group and the control group DPC,the over-expression group and the control group DPC.and to verified the effect of Annexin A2 on proliferation and apoptosis of DPC.The protein which can interact with Annexin A2 was detected by Co-immunoprecipitation to study the signaling pathway of Annexin A2 in DPC.Results: 1.The apoptosis rate and the apoptotic factor Bcl-2 of the passage 3DPC was significantly lower than the passage 10 DPC.On the contrary,the proliferation and the proliferative factor PCNA significantly higher than that of the passage 10 DPC.2.After transfected with the adenovirus,it was observed that the transfection efficiency of DPC was more than 90%.q RT-PCR suggested that the expression of Annexin A2 m RNA in the passage 3 DPC was reduced after transfected with ADV1-ANXA2 which knocking down Annexin A2.After transfected with ADV6-ANXA2 which over-expressing Annexin A2,the expression level of Annexin A2 m RNA in the passage 10 DPC was increased.3.After the passage 3 DPC were transfected with ADV1-ANXA2 which knocking down Annexin A2,the Annexin A2 and proliferation factor PCNA in the control group were significantly higher than the knock-down group,and the apoptotic factor Bcl-2 was significantly lower than the knockdown group.After the passage 10 DPC were transfected with ADV6-ANXA2 which over-expressing Annexin A2,the Annexin A2 and apoptotic factor Bcl-2 in the over-expression group were significantly higher than the control group,and the proliferation factor PCNA was significantly lower than the control group.4.After co-immunoprecipitation of the passage 2 DPC,the expression of?-catenin,Wnt5 a,and Wnt1 proteins can be seen on the NC membrane by Western Blotting.Conclusion: Annexin A2 can promote proliferation of DPC in aggregative growth and inhibit proliferation of DPC in non-aggregative growth.There is an interaction between Annexin A2 and ?-catenin,Wnt5 a and Wnt1 proteins,which indicates that Annexin A2 regulates DPC through Wnt signaling pathway.