Notch信号通路抑制剂DAPT对急性白血病细胞增殖及凋亡的作用

Effects of Notch Signaling Pathway Inhibitor DAPT on Proliferation and Apoptosis of Acute Leukemia Cells

目的:探讨Notch信号通路抑制剂DAPT对急性白血病细胞增殖及凋亡的作用。方法:选取2017年4月-2018年3月广东医科大学附属医院未化疗白血病患者20例为研究对象,根据形态-免疫-细胞遗传学-分子生物学(MICM)诊断分类,将研究对象骨髓样本中的单个核细胞分离并分为急性淋巴细胞白血病(acute lymphocyte leukemia,ALL)和急性髓系白血病(acute myeloid leukemia,AML),ALL与AML经纯化扩增后分别随机分为实验组及对照组,实验组均予以通路抑制剂DAPT 1 μmol/L共培养,对照组予以等量二甲基亚砜DMSO共培养,共培养72 h后计算各组细胞增殖抑制率,同时应用Annexin V-FIFC/PI检测细胞凋亡情况。结果:ALL中实验组的增殖抑制OD值低于对照组,差异有统计学意义(P<0.000 1);AML中两组的增殖抑制OD值比较,差异无统计学意义(P>0.05)。另外AML予DAPT干预72 h后对细胞进行计数,其中实验组细胞个数为8.6×10^5/mL,对照组细胞个数为9.2×10^5/mL,两组细胞数量比较,差异无统计学意义(P>0.05)。AML中实验组细胞凋亡率与对照组比较,差异无统计学意义(P>0.05)。ALL的实验组在培养72 h后细胞计数为5.2×10^5/mL,而对照组细胞计数9.0×10^5/mL,实验组的细胞计数明显低于对照组,差异有统计学意义(P<0.05)。ALL中实验组细胞凋亡率为(15.78±0.48)%,高于对照组的(6.25±0.49)%,差异有统计学意义(P<0.001)。结论:Notch信号通路抑制剂对ALL白血病细胞的增殖及凋亡起到重要的作用,而对AML无明显影响。

Objective:To investigate the effect of Notch signaling pathway inhibitor DAPT on proliferation and apoptosis of acute leukemia cells.Method:A total of 20 non-chemotherapy leukemia patients in affiliated hospital of Guangdong Medical University from April 2017 to March 2018 were selected as the research objects,according to morphoid-immunocytogenetic-molecular biology (MICM) diagnostic classification,the mononuclear cells in the bone marrow samples of research objects were isolated and divided into acute lymphocyte leukemia (ALL) and acute myeloid leukemia (AML).After the purification and amplification of ALL and AML,they were randomly divided into the experimental group and the control group,the experimental group was co-cultured with the pathway inhibitor DAPT 1 μmol/L,and the control group was co-cultured with the same amount of dimethylsulfoxide DMSO,after co-culture for 72 hours,the inhibition rate of cell proliferation in each group was calculated.Result:In ALL,the proliferation inhibition OD value of the experimental group was lower than that of the control group,the difference was statistically significant (P<0.000 1);in AML,there was no statistically significant difference in proliferation inhibition OD values between the two groups (P>0.05).In addition,AML cells were counted after DAPT intervention for 72 h,the number of cells in the experimental group was 8.6×10^5/mL,and 9.2×10^5/mL in the control group,there was no significant difference in cell number between the two groups (P>0.05).There was no statistically significant difference in apoptosis rate between the experimental group and the control group in AML (P<0.05).In ALL,after 72 h of culture,the cell count of the experimental group was 5.2×10^5/mL,while 9.0×10^5/mL of the control group,the cell count of the experimental group was significantly lower than that of the control group,with statistically significant difference (P<0.05).In ALL,the apoptosis rate of the experimental group was (15.78±0.48)%,higher than (6.25±0.49)% of the control group,the difference was statistically significant (P<0.001).Conclusion:Notch signaling pathway inhibitor plays an important role in the proliferation and apoptosis of ALL leukemia cells,but has no significant effect on AML.