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

作　　者：曹秀丽 黄佩 马金花 黄茂林 官亚宁 陈艳 何志旭 Cao Xiuli;Huang Pei;Ma Jinhua;Huang Maolin;Guan Yaning;Chen Yan;He Zhixu(Department of Pediatrics,Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563006,China;Department of Pediatrics,Guizhou Children's Hospital,Zunyi Guizhou 563006,China;Collaborative Innovation Center for Tissue Injury Repair and Regenerative Medicine of Zunyi Medical University,Zunyi Guizhou 563003,China)

机构地区：[1]遵义医科大学附属医院小儿内科,贵州遵义563006 [2]贵州省儿童医院小儿内科,贵州遵义563006 [3]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

出　　处：《遵义医科大学学报》2024年第4期376-383,共8页Journal of Zunyi Medical University

基　　金：省部共建协同创新中心项目[NO:教科技厅函(2020)39];贵州省科技计划项目[NO:黔科合平台人才-CXTD(2021)010];遵义市科技计划项目[NO:遵市科合HZ字(2021)186]。

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、2μmol/L)、ULK1抑制剂MRT68921(0、1.2、2μmol/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。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 activator 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 mitochondrial 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 inhibitor 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 membrane 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 decreased,but ULK1 activator had no significant effect on the CEM-C7 cells.Conclusion ULK1 may affect the apoptosis 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 

分 类 号：R733.71[医药卫生—肿瘤]