胰岛素通过调控PI3K和Erk通路促进人白血病细胞株K562增殖和迁移

目的:观察胰岛素对白血病细胞株K562中PI3K及Erk信号通路的影响及其对K562细胞增殖和迁移的影响?方法:分别采用0?25?50?100和200 nmol/L胰岛素处理K562细胞48 h及200 nmol/L胰岛素处理K562细胞0?12?24和48 h?采用Western blot方法检测K562细胞中PI3K及Erk信号通路的活化情况;采用MTT方法检测胰岛素对K562细胞增殖的影响;采用伤口愈合实验检测胰岛素对K562细胞迁移能力的影响;采用PI3K及Erk信号通路抑制剂LY294002及U0126处理K562细胞,观察PI3K及Erk信号通路在胰岛素促K562细胞增殖和迁移中的作用?结果:同0 nmol/L胰岛素处理组相比,25?50?100?200 nmol/L的胰岛素均可显著上调K562细胞中PI3K及Erk的磷酸化水平,加强K562细胞的增殖能力(P < 0.01)?同0 h组相比,200 nmol/L的胰岛素处理K562细胞12?24和48 h,亦可显著上调K562细胞中PI3K及Erk的磷酸化水平,加强K562细胞的增殖能力(P < 0.01)?同时,同对照组相比,200 nmol/L胰岛素处理48 h后,K562细胞迁移能力均显著增强(P < 0.01)?采用PI3K及Erk信号通路抑制剂LY294002及U0126预处理K562细胞后,胰岛素促K562细胞增殖和迁移的能力显著降低(P < 0.01)?结论:胰岛素可通过激活K562细胞中的PI3K及Erk信号通路,进而增强K562细胞的增殖和迁移能力?

Insulin promotes K562 cells proliferation and migration through PI3K and Erk signaling pathway

Objective:To observe the effect of insulin on PI3K and Erk signaling pathway and its regulation of proliferation and migration in K562 cells. Methods: K562 cells were treated with insulin with different concentrations (0, 25, 50, 100 and 200 nmol/L) for 48 h, and 200 nmol/L insulin for 0, 12, 24 and 48 h. Then, the phosphorylation levels of PI3K and Erk were detected by Western blot, and the proliferation ability of K562 cells was detected by MTT. Effect of insulin on migration ability of K562 cells was detected by wound healing assay. K562 cells were pretreated with PI3K and Erk signaling pathway inhibitors LY294002 and U0126 to observe the effect of PI3K and Erk signaling pathway on insulin induced proliferation and migration in K562 cells. Results: Compared with the 0 nmol/L insulin treated group, 25, 50, 100 and 200 nmol/L insulin treated groups had significantly increased phosphorylation levels of PI3K and Erk and promoted proliferation in K562 cells (P < 0.01). Compared with the 0 h insulin treated group, 200 nmol/L insulin treated for 12, 24 and 48 h groups also had significantly increased phosphorylation levels of PI3K and Erk and promoted proliferation in K562 cells (P < 0.01). After treated with 200 nmol/L insulin by 48 h, the migration ability of K562 cells were significantly increased (P < 0.01). After blocked PI3K and Erk signaling pathway by LY294002 and U0126, the ability of insulin in induced proliferation and migration in K562 cells were significantly decreased (P < 0.01). Conclusion: Through actives PI3K and Erk signaling pathway, insulin can promote proliferation and migration of K562 cells.