硫柳汞对白血病细胞株凋亡及自噬调控的研究  

Effects of Thiomersal on Apoptosis and Autophagy of Leukemia Cell Lines

作　　者：王莹[1,2] 姚浩 李转丽 杨柯 白海 WANG Ying;YAO Hao;LI Zhuan-Li;YANG Ke;BAI Hai(Deparment of Hematology,The 940th Hospital of Joint Logistics Support Force of Chinese People′s Liberation Army,Lanzhou 730050,Gansu Province,China;Deparment of Hematology,The General Hospital of Western Theater Command,PLA,Chengdu 610083,Sichuan Province,China)

机构地区：[1]中国人民解放军联勤保障部队第940医院血液科,甘肃兰州730050 [2]中国人民解放军西部战区总医院血液科,四川成都610083

出　　处：《中国实验血液学杂志》2022年第6期1655-1660,共6页Journal of Experimental Hematology

摘　　要：目的:探讨不同浓度硫柳汞对人白血病细胞株U937、CEM-C1、BALL-1凋亡及自噬调控的影响。方法:应用CCK-8法检测硫柳汞对U937、CEM-C1、BALL-1细胞增殖抑制作用,Annexin V FITC/PI双染流式细胞术检测其凋亡率,Western blot法检测硫柳汞对细胞自噬信号通路PI3K/Akt/mTOR以及PI3K、Akt、mT OR、p-mT OR、Caspase-3、LC3-II蛋白表达的影响。结果:在24和48 h内,硫柳汞对U937、CEM-C1、BALL-1细胞系具有增殖抑制作用且呈时间、剂量依赖性(r_(24 h)=0.295,r_(24 h)=0.452,r_(24 h)=0.103;r_(48 h)=0.821,r_(48 h)=0.665,r_(48 h)=0.821),而72 h未见明显的时间、剂量依赖性。硫柳汞处理U937、CEM-C1和BALL-1细胞48 h,其凋亡率均呈剂量依赖性升高(r=0.819,r=0.763,r=0.835)。硫柳汞作用U937、CEM-C1、BALL-1细胞48 h,细胞PI3K、AKT、mT OR、r-mT OR蛋白表达呈浓度依赖性下调,其中U937细胞值分别为-0.975、-0.899、-0.925、-0.915,CEM-C1细胞r p值分别为-0.960、-0.920、-0.861、r=-0.927,BALL-1细胞r值分别为-0.939、-0.911、-0.896、-0.926,表明硫柳汞诱导U937、CEM-C1、BALL-1细胞凋亡可能是因为抑制了PI3K/AKT/mTOR通路。硫柳汞通过Caspase-3途径促进U937、CEM-C1、BALL-1细胞凋亡,Caspase-3、LC3-II表达呈剂量依赖性上调(r=0.976,r=0.914;r=0.976,r=0.986;r=0.961,r=0.974)。结论:硫柳汞能够抑制U937、CEM-C1、BALL-1细胞增殖并促进其凋亡,一定浓度的硫柳汞可下调U937、CEM-C1、BALL-1细胞PI3K/Akt/mTOR通路相关蛋白PI3K、AKT、mT OR、p-mT OR的表达,同时通过下调PI3K/Akt/mTOR通路激活U937、CEM-C1、BALL-1细胞自噬、诱导凋亡。Objective:To investigate the effects of different concentrations of thiomersal on apoptosis and autophagy regulation of human leukemia cell lines U937,CEM-C1 and BALL-1.Methods:The inhibitory effect of thiomersal on the proliferation of U937,CEM-C1 and BALL-1 cells was detected by CCK-8 assay.Annexin V-FITC/PI double staining flow cytometry was used to detect the apoptosis rate.Western blot was used to detect the effects of thiomersal on autophagy signaling pathway and the expression of PI3 K,Akt,mTOR,p-mTOR,caspase-3 and LC3-II proteins.Results:Within 24 and 48 hours,thiomersal inhibited the proliferation of U937,CEM-C1 and BALL-1 cell lines in a time and dose-dependent manner(r_(24 h)=0.295,r_(24 h)=0.452,r_(24 h)=0.103;r_(48 h)=0.821,r_(48 h)=0.665,r_(48 h)=0.821),but no significant time and dose-dependent effect was observed at 72 hours.After 48 hours treatment of thiomersal,the apoptosis rate of U937,CEM-C1 and BALL-1 cells increased in a dose-dependent manner(r=0.819,r=0.763,r=0.835).After 48 hours treatment of thiomersal,the expression levels of PI3 K,Akt,mTOR and p-mTOR protein in U937,CEM-C1 and BALL-1 cells decreased in a concentration-dependent manner,the R value of U937 cells was-0.975,-0.899,-0.925 and-0.915,respectively,that of CEM-Cl cells was-0.960,-0.920,-0.861 and-0.927,and that of BALL-1 cells was-0.939,-0.911,-0.896 and-0.926,.which suggested that thiomersal-induced apoptosis of U937,CEM-C1 and BALL-1 cells might be due to the inhibition of PI3 K/Akt/mTOR pathway.Thiomersal promoted the apoptosis of U937,CEM-Cl and BALL-1 cells via caspase-3 pathway,and the expressions of caspase-3 and LC3-II were up-regulated in a dose-dependent manner(r=0.976,r=0.914;r=0.976,r=0.986;r=0.961,r=0.974).Conclusions:Thiomersal can inhibit the proliferation and promote the apoptosis of U937,CEM-C1 and BALL-1 cells.A certain concentration of thiomersal can down-regulate the expression of PI3 K/Akt/mTOR pathway related proteins PI3 K,Akt,mTOR and p-mTOR in U937,CEM-C1 and BALL-1 cells,and activate autophagy and apop

关 键 词：硫柳汞 白血病 PI3K/Akt/mTOR通路 自噬 凋亡 

分 类 号：R733[医药卫生—肿瘤]