白细胞介素⁃33对急性髓系白血病细胞株HL⁃60、NB4凋亡和周期的调控作用及机制研究

Regulation and mechanism of IL⁃33 on apoptosis and cell cycle of acute myeloid leukemia cell lines HL⁃60 and NB4

目的探讨白细胞介素⁃33(interleukin⁃33,IL⁃33)对急性髓系白血病细胞凋亡的影响及其可能的机制。方法急性髓系白血病细胞株HL⁃60、NB4经外源性IL⁃33以及p38丝裂原活化蛋白激酶(p38 mitogen⁃activated protein kinase,p38 MAPK)特异抑制剂(SB203580)处理72 h后,采用Annexin V/DAPI双染色流式细胞术检测细胞凋亡率,细胞周期检测试剂盒检测细胞周期,Western blot检测细胞周期蛋白依赖性激酶1(cyclin⁃dependent kinases 1,CDK1)蛋白表达以及p38 MAPK的磷酸化水平。结果与对照组相比,IL⁃33组的HL⁃60和NB4细胞经IL⁃33作用后,细胞凋亡水平降低(P<0.05),细胞周期中S期比例升高(P<0.001);而SB+IL⁃33组细胞经SB203580和IL⁃33联合处理后,细胞凋亡水平高于IL⁃33组(P<0.05),S期细胞的百分比低于IL⁃33组(P<0.05);Western blot检测结果表明,与对照组相比,IL⁃33组细胞的p38 MAPK磷酸化(p⁃p38 MAPK)水平和CDK1蛋白表达水平均显著增高(均P<0.05),而SB+IL⁃33组的p⁃p38 MAPK水平以及CDK1水平较IL⁃33组显著下降(均P<0.05)。结论IL⁃33可通过激活p38 MAPK,抑制急性髓系白血病细胞株HL⁃60、NB4细胞的凋亡,抑制细胞周期停滞。

Objective To investigate the regulation and mechanism of interleukin⁃33(IL⁃33)on the apoptosis and cell cycle of acute myeloid leukemia(AML)cells HL⁃60 and NB4.Methods AML cell lines HL⁃60 and NB4 were treated with IL⁃33 and p38 mitogen⁃activated protein kinase(p38 MAPK)inhibitor(SB203580)for 72 h.The cell apoptosis rate was detected by Annexin V/DAPI double staining flow cytometry and the cell cycle was detected by cell cycle detection kit.The expression levels of CDK1 protein and p38 MAPK phosphorylation were detected by Western blot.Results Compared with the control group,the apoptosis level of HL⁃60 and NB4 cells in IL⁃33 group was reduced after IL⁃33 treatment(P<0.05),the proportion of S phase in cell cycle increased(P<0.001);while the apoptosis level of SB+IL⁃33 group was higher than that of IL⁃33 group after combined treatment with SB203580 and IL⁃33(P<0.05),and the percentage of S phase cells was lower than that of IL⁃33 group(P<0.05).Western blot results showed that the phosphorylation level of p38 MAPK(p⁃p38 MAPK)and the expression level of CDK1 protein were significantly increased in IL⁃33 group,compared with the control group(all P<0.05).The expression levels of p⁃p38 MAPK and CDK1 in SB+IL⁃33 group were significantly lower than those in IL⁃33 group(all P<0.05).Conclusions IL⁃33 can inhibit apoptosis and cell cycle of AML cell lines HL⁃60 and NB4 by activating p38 MAPK.