小分子化合物TIC10对白血病细胞的抑制作用

Inhibitory Effect of Small Molecule Compound TIC10 on Leukemia

目的:研究小分子化合物TIC10对多种白血病细胞的抑制作用及其对MLL-AF9诱发的急性髓系白血病的干预效果。方法:采用MOLM-13、MV4-11、THP-1、Kasumi-1、KG-1和K562细胞,通过CCK-8和细胞计数检测TIC10对白血病细胞增殖的影响。BrdU和DAPI染色的流式细胞术检测TIC10对白血病细胞周期的影响。Annexin V和PI染色检测TIC10对白血病细胞凋亡的影响。Western blot检测TIC10对白血病细胞凋亡关键蛋白Caspase3表达的影响。构建小鼠MLL-AF9白血病模型以检测TIC10对急性髓系白血病在体进展的干预效果。结果:TIC10处理均显著抑制MOLM-13、MV4-11、THP-1和Kasumi-1细胞的增殖(P<0.05)。TIC10处理使MV4-11及Kasumi-1细胞中的BrdU掺入显著减少(P<0.01),表明TIC10阻碍白血病细胞进入S期。凋亡实验结果显示,TIC10能显著诱导白血病细胞凋亡(P<0.01)。免疫印迹结果表明,TIC10促进多种白血病细胞中Cleaved-Caspase3蛋白水平的增加(P<0.05)。动物实验结果证实TIC10能有效干预小鼠体内白血病的进展。结论:TIC10能有效抑制白血病细胞的增殖并诱导细胞凋亡,对MLL-AF9诱发的急性髓系白血病有一定干预效果,TIC10有望作为白血病治疗的潜在候选药物。

Objective:To investigate the inhibitory effect of small molecule compound TIC10 on multiple leukemia cells and its intervention effect on acute myeloid leukemia(AML)induced by MLL-AF9.Methods:Leukemia cells(MOLM-13,MV4-11,THP-1,Kasumi-1,KG-1 and K562)were used to detect the inhibitory effect of TIC10 on proliferation of leukemia cells through CCK-8 and cell count.The effect of TIC10 on the cell-cycle of leukemia cells was detected by flow cytometry with BrdU and DAPI staining.Apoptosis was detected by Annexin V and PI staining.Western blot was used to detect the effect of TIC10 on the expression of caspase3,a key apoptosis-associated protein.The intervention effect of TIC10 on the progression of leukemia in vivo was detected by constructing a mouse MLL-AF9 leukemia model.Results:TIC10 treatment significantly inhibited the proliferation of MOLM-13,MV4-11,THP-1,and Kasumi-1 cells(P<0.05).TIC10 treatment substantially reduced the incorporation of BrdU in MV4-11 and Kasumi-1 cells(P<0.01),suggesting that TIC10 inhibited S-phase entry of leukemia cells.Apoptosis assay showed that TIC10 obviously induced apoptosis of leukemia cells(P<0.01).Western blot results showed that TIC10 treatment induced an elevated cleaved-caspase3 protein level in leukemia cells(P<0.05).The results of animal experiments proved that TIC10 effectively interfered with the progression of AML in vivo.Conclusion:TIC10 can effectively inhibite the proliferation of leukemia cells and induce apoptosis,and have a certain intervention effect on AML induced by MLL-AF9,indicating that TIC10 as a potential candidate drug for the treatment of leukemia.