IL-15对HaCaT细胞增殖的影响及机制研究

Study on the impact of IL-15 on proliferation of HaCaT cells

目的观察白介素-15(IL-15)对人皮肤角质形成细胞(HaCaT)增殖的影响及其可能机制。方法培养HaCaT细胞,不同剂量IL-15刺激不同时间后,MTT法分析IL-15对HaCaT细胞增殖的影响。Western blot分析IL-15在HaCaT细胞中激活的信号通路:丝裂原蛋白活化激酶-胞外信号调节激酶(MAPKs-ERK1/2)和磷脂酰肌醇激酶-丝氨酸/苏氨酸激酶(PI3K-AKT)。在IL-15刺激HaCaT细胞前1h,分别加入MAPKs-ERK1/2和PI3K-AKT信号传导特异性抑制剂(PD98059和LY294002)以阻断IL-15激活的相关信号通路,分析阻断剂对IL-15调控HaCaT细胞增殖的影响。结果 MTT分析结果显示:IL-15显著促进HaCaT细胞增殖,且具有时间及剂量依赖性;信号通路分析揭示IL-15能够增加pERK1/2及pAKT的水平;使用阻断剂的研究显示IL-15部分依赖MAPKs-ERK1/2和PI3K-AKT途径发挥其促进增殖作用。结论 IL-15部分依赖MAPKs-ERK1/2和PI3K-AKT信号途径促进HaCaT细胞增殖。

Objective To explore the impact of IL-15 on proliferation of HaCaT cells. Methods HaCaT cells were cultured and treated with various concentration of IL-15 for different time. MTT method was used to analyze the proliferation of HaCaT cells. Furthernore, we analyzed signaling pathways induced by IL-15 in HaCaT cells by Western-Blot, and then used the specific inhibitors to selectively inhibit ERK and AKT phosphorylation to reveal the relationship between the affect of IL-15 on the proliferation and these signaling pathways. Results In HaCaT cells, IL-15 promotes the proliferation of HaCaT cells at a dose-dependent and time-dependent ways. Mechanistic studies show that IL-15 increased the phosphorylation of ERK1/2 and AKT, and the specific inhibitors of the phosphorylation of ERK1/2 （ PD98059 ） or AKT （ LY2940002 ） could antagonize the effect of IL-15 on the proliferation of HaCaT cells. Conclusion The IL-15 partially depends on the MAPKs-ERK1/2 and PI3K-AKT signaling pathways to regulate the proliferation in HaCaT cells.