血清饥饿对人白血病细胞株THP-1细胞增殖、细胞死亡和细胞自噬的影响  

Effects of serum deprivation on cell proliferation,cell death and autophagy in human leukemia cell line THP-1

作　　者：高雪[1] 郑晓琦 刘为忠[1] 刘德胜[1] 张静[1] 潘效红[1] 

机构地区：[1]滨州医学院药学院细胞工程研究室,烟台264003

出　　处：《山西医科大学学报》2017年第10期1003-1007,共5页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(81573412;31270082);山东省自然科学基金资助项目(ZR2014JL055;ZR2013HM042);滨州医学院科研启动基金资助项目(BY2015KYQD03)

摘　　要：目的本实验通过血清饥饿处理人急性单核细胞白血病细胞株THP-1,探究血清饥饿对细胞增殖、细胞死亡和细胞自噬的影响及其机制。方法实验分为4组:0%FBS组、1%FBS组、5%FBS组和10%FBS组(对照组),血清饥饿时长为24 h和48 h。通过MTT法检测细胞增殖,台盼蓝拒染法检测细胞死亡,流式细胞术测定吖啶橙(AO)的荧光强度,免疫印迹法检测自噬相关蛋白Atg5、Atg7和LC3Ⅱ表达。结果与对照组相比,不同程度血清饥饿处理24 h和48 h均可以显著抑制THP-1细胞增殖(P<0.05),细胞增殖呈现一定的血清浓度依赖性和饥饿时间依赖性。血清饥饿处理24 h和48 h可以显著增加细胞死亡率(P<0.05),并增加细胞内酸性自噬体囊泡的形成。免疫印迹显示,血清饥饿显著性增强自噬相关蛋白Atg5,Atg7和LC3Ⅱ的表达(P<0.05)。结论血清饥饿可降低THP-1细胞增殖,增加细胞死亡率和自噬。Objective To investigate the effects of serum deprivation on cell proliferation,cell death and autophagy in human monocytic leukemia cell line THP-1. Methods Experiments were divided into four groups： 0% FBS group,1% FBS group,5% FBS and10% FBS group（ control group）,and cells were starved for 24 h and 48 h. Cell proliferation was performed by MTT assay. Cell death was tested by trypan blue exclusion assay. Fluorescence intensity of acridine orange（ AO） was examined by a flow cytometer. Western blot was performed for investigating the expression of autophagy-related genes at the protein level. Results The serum deprivation significantly inhibited cell proliferation in a concentration-dependent and time-dependent manner after FBS treatment（ P 〈 0. 05）. And serum deprivation significantly increased cell death and the fluorescence intensity of acid autolysosome（ P 〈 0. 05）. Serum deprivation signifi cantly increased the expression of Atg5,Atg7 and LC3Ⅱ（ P 〈 0. 05）. Conclusion Serum deprivation can inhibit the cell proliferation,induce cell death and autophagy of THP-1 cells.

关 键 词：THP-1细胞 血清饥饿 细胞增殖 细胞死亡 细胞自噬 

分 类 号：Q279[生物学—细胞生物学]