法舒地尔对HL-60细胞增殖、凋亡、分化及侵袭作用的影响

摘要

Objective:To investigate the effect of Rho kinase inhibitor fasudil on proliferation,differentiation,apoptosis and migration of HL-60 cells.Methods:Cell growth curve was analyzed by trypan blue staining,and the inhibitory effect of fasudil on the proliferation of HL-60 cells MTT assay.The differentiation of HL-60 cells exposed to different concentrations of fasudil was evaluated by expression of CD 11b using flow cytometry and NBT reduction test.Cell apoptosis rate detected with AnnexinV/FADD double staining by flow cytometry.Trans well chamber in vitro tumor cell migration model was observed the effects of fasudil on leukemic cell migration.Results:Fasudil in-hibited growth of HL-60 cellsin a time-and dose-dependent manner.When HL-60cells was treated by fasudil at 12h,24h,48h and 72h,the median inhibitory concentration(IC50)were(89.65 ± 12.45)μmol/L,(69.21 ± 15.48)μmol/L,(65.29 ± 20.31)μmol/L and(58.54 ± 10.02)μmol/L,respectively.Fasudil induced apoptosis of HL-60 cells in time-and-dose effect.Fasudil could upregulate the expression of CD11b of HL-60 cells and increased position of NBT in HL-60 cells.Fasudil reduced Significantly fewer HL-60 cells were migrated into the lower compartment.Conclu-sion:Fasudil can inhibit the proliferation of leukemic cells and induce HL-60 cells apoptosis,promote the differentia-tion of leukemia cells,can inhibit leukemic cells distant metastasis.%目的:探讨ROCK 酶抑制剂法舒地尔对 HL-60细胞增殖、凋亡、分化以及侵袭作用的影响.方法:用台盼蓝拒染实验绘制法舒地尔作用下,HL-60细胞的生长曲线;M T T 法检测法舒地尔对HL-60细胞增殖抑制.应用流式细胞术CD11b抗原表达,硝基四唑氮蓝(NBT)还原实验分析法舒地尔对HL-60细胞分化影响.采用AnnexinV/PI双染试剂盒上流式细胞仪测定细胞凋亡.利用Transwell穿孔板法观察法舒地尔对HL-60细胞侵袭力的影响.结果:法舒地尔抑制HL-60细胞呈浓度和时间依赖性、法舒地尔作用于 H L-60细胞12、24、48、72 h的半数抑制浓度(IC50)分别为(89.65 ± 12.45)μmol/L、(69.21 ± 15.48)μmol/L、(65.29 ± 20.31)μmol/L 以及(58.54 ± 10.02)μmol/L.法舒地尔诱导 H L-60细胞凋亡呈时间和剂量依赖性.法舒地尔可增加HL-60细胞后硝基四唑氮蓝阳性率,上调CD11b单核分化抗原的表达.法舒地尔明显抑制 HL-60细胞穿透T ransw ell小室,侵袭到穿孔膜外.结论:法舒地尔有抑制HL-60细胞增殖、诱导凋亡、促进分化及抑制侵袭等多种抗白血病作用.