IL-10on The Proliferation And Differentiation Of HaCaT Cells

Objective To explore the effect of the IL-10on the proliferation and differentiation ofHaCaT cells, and then uncover the mechanism.Method HaCaT cells were cultured and treated with various concentration of theIL-10for different time. MTT and FACS methods were used to analyze the the effect ofIL-10on the proliferation of HaCaT cells. Though treated with various concentration ofthe IL-10for different time, the makers (the layer of granule Keratin1, the layer ofbasis Keratin5and the layer of thorn Involucrin) of HaCaT cells differentiation inducedby the certain concentration of CaCl2were detected by Western Blot; Furthermore,Western Blot was used to analyzed signaling pathway activated by IL-10, includingmitogen-activation protein kinase-extracellular signal-regulatedkinase(MAPKs-ERK1/2) and phosphatidylinositol kinase-serine/threoninekinases(PI3K-AKT). Before an hour HaCaT cells were treated with the IL-10we usedthe specific inhibitors(PD98059and LY2940002) selectively inhibit ERK1/2and AKTphosphorylation to reveal the relationship between the affect of IL-10on thedifferentiation and these relative signaling pathways.Results The results of MTT and FACS highlighted that the IL-10didn’t effect theproliferation of HaCaT cells; However, IL-10promoted the expression of differentiationmakers (K1, INV) in HaCaT cells induced by CaCl2at protein levels; Mechanisticstudies show that the IL-10increased the phosphorylation of ERK1/2and AKT, andthe specific inhibitors of the phosphorylation of ERK1/2(PD98059) or AKT(LY2940002) could antagonize the effect of the IL-10on the phosphorylation of HaCaTcells. The studies of the inhibitors of the phosphorylation of ERK1/2(PD98059) andAKT (LY294002) showed the effect of IL-10on the differentiation was partly dependent of the MAPKs-ERK1/2and PI3K-AKT signaling pathway.Conclusion IL-10partly depends on the PI3K-AKT and MAPKs-ERK1/2signalingpathways to regulate the differentiation in HaCaT cells.