LY294002对多发性骨髓瘤细胞增殖的抑制作用及机制研究

摘要

目的:探讨2-(4-吗啉基)-8-苯基4氢-1-苯并吡喃4酮(LY294002)对多发性骨髓瘤细胞株U266的增殖抑制作用及其机制.方法:用不同浓度(0、5、10和20 μmol/L) LY294002处理U266细胞24、48和72 h后,MTT法检测细胞活力,吖啶橙染色法检测细胞形态,流式细胞术检测细胞周期,Western blot检测细胞中B淋巴细胞瘤-2(BCL-2)、BCL-2相关X蛋白(BAX),细胞周期蛋白D1(Cyclin D1),Cyclin E表达及磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)通路的激活状况.结果:0、5、10和20 μmol/L的LY294002处理U266细胞24、48和72 h后,能显著提高细胞增殖抑制率,并具有时间及剂量依赖性(r =0.408,r =0.485).5,10,20 μmol/L的LY294002处理细胞48 h,能使细胞核呈现致密浓染,细胞周期明显阻滞在G1期(P≤0.01),显著下调BCL-2,Cyclin D1,Cyclin E,PI3K及p-AKT表达(P≤0.01),明显上调BAX表达(P≤0.01).结论:LY294002能显著抑制多发性骨髓瘤细胞U266增殖,其作用机制可能与抑制PI3 K/AKT信号通路的激活有关.%Objective:To explore the inhibitory effect of 2-(4-morpholine)-8-phenyl-4 hydrogen-1-benzo-4 ketone (LY294002) on proliferation of multiple myeloma cell U266 and its mechanism.Methods:U266 cells were cultured with 0,5,10,20 μmol/L LY294002 for 24,48 and 72 h,the cell viability was measured by MTT method,cell morphology was observed by acridine orange staining,cell cycle was measured by flow cytometry.The expressions of B-cell lymphoma-2 (BCL-2),BCL-2-associated X protein (BAX),Cyclin D1,Cyclin E and activation of phosphatidyl inositol 3 kinase (PI3K)/protein kinase B (AKT) signal pathway were measured by Western blot.Results:U266 cell viability was reduced in time-and dose-dependent manner after treatment with 5,10,20 μmol/L of LY294002 for 24,48,72 h.The 5,10,20 μmol/L LY294002 leaded to cell nucleus dense and thick,and the cell cycle arrested in the G1 phase (P ≤0.01).The expressions of BCL-2,Cyclin D1,Cyclin E,PI3K and p-AKT were downregulated (P ≤0.01),and the expression of BAX up-regulated (P ≤ 0.01).Conclusion:LY294002 can inhibit U266 cell proliferation via suppresion of activation of PI3K/AKT signal pathway.