激酶ULK1对急性T淋巴细胞白血病细胞凋亡作用的研究
Effect of kinase ULK1 on apoptosis of acute T lymphocytic leukemia cells
曹秀丽 1黄佩 1马金花 1黄茂林 1官亚宁 1陈艳 1何志旭1
作者信息
- 1. 遵义医科大学附属医院小儿内科,贵州遵义 563006;贵州省儿童医院小儿内科,贵州遵义 563006;遵义医科大学组织损伤修复与再生医学省部共建协同创新中心,贵州遵义 563003
- 折叠
摘要
目的 探讨ULK1对急性T淋巴细胞白血病细胞株CEM-C7凋亡的影响及与AMPKα/ULK1轴之间的关系.方法 通过予不同浓度 ULK1 激活剂 LYN-1604(0、1.2、2 μmol/L)、ULK1 抑制剂 MRT68921(0、1.2、2 μmol/L)干预 CEM-C7 细胞24 h,流式细胞术检测各药物浓度对CEM-C7细胞株凋亡率的影响,筛选凋亡最明显的药物浓度进行后续实验,Edu及Soft agar实验检测其对CEM-C7细胞株增殖的影响,流式细胞术检测其对CEM-C7细胞株线粒体膜电位的影响,透射电镜观察其对CEM-C7细胞细胞核及线粒体超微结构变化的影响,Western blot检测激活和抑制ULK1后ULK1、p-ULK1(Ser317)、Caspase3、Cleaved-caspase3、Bcl-2、Bax、PCNA、AMPKα、p-AMPKα(Ser172)的表达情况.结果 ULK1 抑制剂及激活剂分别干预CEM-C7细胞24 h以后,与对照组相比,ULK1抑制剂可明显促进CEM-C7细胞发生凋亡,抑制CEM-C7细胞增殖,使细胞线粒体的膜电位明显下降,差异均具有统计学意义(P＜0.05);透射电镜可观察到明显的细胞凋亡现象;p-ULK1(Ser317)、p-AMPKα(Ser172)的蛋白水平表达下降,而ULK1激活剂对CEM-C7细胞株无明显影响.结论 ULK1可能通过AMPKα/ULK1轴影响其对CEM-C7细胞的凋亡功能.
Abstract
Objective To investigate the effect of ULK1 on apoptosis of acute T lymphocytic leukemia cell line CEM-C7 and its relationship with AMPKα/ULK1 axis.Methods The CEM-C7 cells were treated with ULK1 ac-tivator LYN 1604(0,1.2,and 2 μmol/L),and ULK1 inhibitor MRT68921(0,1.2,and 2 μmol/L)for 24 h.The apoptosis rate of the CEM-C7 cells was detected by flow cytometry.The drug concentration with the most obvious apoptosis effect was screened for follow-up experiments.Edu and Soft agar tests were conducted to detect its effect on the proliferation of CEM-C7 cells,flow Bax cytometry was detected to explain its effect on the mito-chondrial membrane potential of CEM-C7 cells,and the changes on the nucleus and mitochondrial ultrastructure of the CEM-C7 cells were observed under transmission electron microscopy.The expressions of ULK1,p-ULK1(Ser317),Caspase3,Cleaved caspase3,Bcl-2,Bax,PCNA,AMPKα,and p-AMPKα(Ser172)were detected by Western blot analysis after activation and inhibition of ULK1.Results After 24 h intervention with ULK1 in-hibitor and activator,compared with the control group,ULK1 inhibitor could significantly promote the apoptosis of CEM-C7 cells in a dose-dependent manner,inhibit the proliferation of CEM-C7 cells,and decrease the mem-brane potential of mitochondria,with statistical significance(P＜0.05).The apoptosis could be observed by transmission electron microscope.The protein levels of p-ULK1(Ser317)and p-AMPKα(Ser172)were de-creased,but ULK1 activator had no significant effect on the CEM-C7 cells.Conclusion ULK1 may affect the ap-optosis of CEM-C7 cells through AMPKα/ULK1 axis.
关键词
UNC-51样激酶1/急性T淋巴细胞白血病/CEM-C7细胞/凋亡Key words
Unc51-like autophagy activating kinase 1/acute T lymphocytic leukemia/CEM-C7 cells/apopto-sis引用本文复制引用
基金项目
省部共建协同创新中心项目(教科技厅函202039)
贵州省科技计划(黔科合平台人才-CXTD2021010)
遵义市科技计划(遵市科合HZ字2021186)
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据