Effects Of Bozhi Glycopiptide On Proliferation And Differentiation Of Keratinocytes Based On MiR-203-related Signaling Pathways

Objective:HaCaT cells were treated with Bozhi glycopeptides before and after transfection with mimic and inhibitor of miR-203.According to the changes of TAp63,SOCS3,STAT3 and keratin expression,the effect of miR-203 on TAp63 and SOCS3,the regulation of miR-203 on STAT3 signaling pathway and the effect of miR-203 on the proliferation and differentiation of KC were clarified.The intervention mechanism of Bozhi glycopeptides on the proliferation and differentiation of KC was explored.Methods:1.Human keratinocyte HaCaT cell line was transfected with mimic and inhibitor of miR-203,and before and after transfection HaCaT cells were treated with Bozhi glycopeptides solution;2.The expression levels of TAp63,SOCS3 and STAT3 mRNA in HaCaT cells before and after transfection and treated with Bozhi glycopeptides were measured by qRT-PCR;3.Western blot was used to detect the expression of TAp63,SOCS3,STAT3,K10 and K16 in HaCaT cells before and after miR-203 mimic and inhibitor transfection and treated with Bozhi glycopeptides.4.Through data processing and analysis,the regulatory effect of miR-203 on KC proliferation and differentiation and the intervention mechanism of Bozhi glycopeptides were clarified.Result:1.The results of qRT-PCR showed that compared with the control group,the expression of TAp63 and SOCS3 mRNA were down-regulated and STAT3 mRNA were up-regulated in miR-203 mimic group,while that in miR-203 inbibitor group was opposite.Compared with the group without Bozhi glycopeptides,Bozhi glycopeptides had no significant effect on the expression of TAp63,SOCS3 and STAT3 mRNA.2.Western blot showed that compared with the control group,the expression of TAp63,SOCS3 and STAT3 protein in miR-203mimic group was down-regulated,and the expression of K10 and K16 was up-regulated.In miR-203 inhibitor group,the expression of TAp63 protein,SOCS3 protein and K16 was up-regulated,while that of STAT3 protein and K10 was down-regulated.3.Compared with miR-203 mimic group,the expression of TAp63 protein and SOCS3 protein was up-regulated in miR-203mimic+Bozhi glycopeptides group.Compared with miR-203 inhibitor group,the expression of TAp63 protein and SOCS3 protein was down-regulated in miR-203 inhibitor+Bozhi glycopeptides group.Conclusion:1.miR-203 in HaCaT cells negatively regulates the expression of TAp63 and SOCS3 by targeting,positively regulates the expression of STAT3 mRNA by JAK/STAT signaling pathway,but the expression of STAT3 protein is not affected by the change of miR-203,suggesting that KC can regulate the expression of STAT3 protein through various mechanisms(signaling pathway)in normal physiological state,so as to keep its expression in a stable state and maintain cell homeostasis.2.The expression of K10 in HaCaT was positively correlated with the level of miR-203,and the expression of K16 was not affected by the change of miR-203.3.Bozhi glycopeptides can intervene the regulation of miR-203 on TAp63 and SOCS3 in both directions,and intervene the effect of miR-203 on STAT3 at the level of transcription and translation,reduce the expression of STAT3 and inhibit the proliferation of KC4.By up-regulating TAp63 and down-regulating SOCS3,STAT3,K10 and K16,Bozhi glycopeptides can regulate the proliferation and differentiation of KC.The results showed that its differentiation promoting effect was stronger than that of proliferation promoting effect.