糖皮质激素对抗血清剥夺诱导的小鼠肝癌细胞凋亡作用及机制研究  

The effects and relevant mechanism of glucocorticoid on the apoptosis of mouse hepatoma cells induced by serum deprivation

作　　者：王贵平 曹明月 李高翔 黄薇 杨楠[2] 刘雁勇 WANG Gui-ping;CAO Ming-yue;LI Gao-xiang;HUANG Wei;YANG Nan;LIU Yan-yong(Medical College,Tibet University,Lhasa 850000,China;Department of Pharmacology,Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences & School of Basic Medicine,Peking Union Medical College,Beijing 100005,China)

机构地区：[1]西藏大学医学院,西藏拉萨850000 [2]中国医学科学院基础医学研究所北京协和医学院基础学院药理学系,北京100005

出　　处：《实用医院临床杂志》2020年第4期1-4,共4页Practical Journal of Clinical Medicine

基　　金：国家科技重大专项(编号:2019ZX09301170);北京市自然科学基金(编号:7172134,7192128);中国医学科学院医学与健康科技创新工程(编号:2016-I2M-3-004);国家自然科学基金(编号:81972688);协和青年科研基金与中央高校基本科研业务费专项基金(编号:3332015113,2017350002)。

摘　　要：目的探讨糖皮质激素对血清剥夺条件下小鼠肝癌细胞系H22凋亡的影响及相关机制。方法血清剥夺诱导H22细胞凋亡的同时分别添加不同浓度的糖皮质激素皮质酮(CORT)以及糖皮质激素受体拮抗剂RU486,将细胞依次分为对照组、血清剥夺组、CORT组(0.25、0.5、1μmol/L)、RU486组(1μmol/L)、CORT+RU486组(1μmol/L CORT+1μmol/LRU486),24 h后流式细胞术染色检测H22细胞凋亡水平变化。同时,在正常血清培养条件下,CORT(1μmol/L)处理H22细胞48 h后提取细胞各组分蛋白进行蛋白质免疫印迹实验,检测糖皮质激素受体(GR)核转位以及Hsp90蛋白表达水平变化。结果与对照组相比,血清剥夺组H22细胞凋亡水平显著升高(P<0.001);CORT处理后细胞凋亡水平与血清剥夺组相比又显著降低(P<0.05),并呈现剂量依赖性;CORT与RU486联合处理后H22细胞凋亡水平与CORT(1μmol/L)组相比又显著升高(P<0.001)。在正常血清培养条件下,CORT处理后H22细胞中GR核转位增加,同时协助GR进行核转位的热休克蛋白(Hsp90)表达水平升高。结论糖皮质激素可以对抗血清剥夺诱导的小鼠肝癌H22细胞凋亡,其机制可能与Hsp90协助下增强的GR核转位有关。Objective To investigate the effects of glucocorticoid on apoptosis of mouse H22 hepatocarcinoma cells induced by serum deprivation and its related mechanism.Methods Mouse H22 cells were exposed to serum deprivation.At the same time,the cells were treated with different concentrations of glucocorticoid corticosterone(CORT)and/or glucocorticoid receptor antagonist RU486.The cells were divided into control group,serum deprivation group,CORT groups(0.25,0.5,and 1μmol/L),RU486 group(1μmol/L)and CORT+RU486 group(CORT 1μmol/L+RU4861μmol/L).After 24 h of treatment,apoptotic level of the cells was detected by flow cytometry staining.Furthermore,the cytosolic and nuclear proteins of the cells were extracted after 48 h treatment with CORT(1μmol/L)under normal culture conditions with serum,and the levels of GR nuclear translocation and Hsp90 protein expression were detected by using western blot analysis.Results Compared with the control group,the apoptosis level of H22 cells in serum deprivation group was significantly increased(P<0.001).Compared with serum deprivation group,CORT treatment significantly decreased the apoptosis level of H22 cells in a dose-dependent manner(P<0.05)and the apoptosis level of H22 cells after co-treatment of CORT and RU486 was significantly increased compared with the CORT group(P<0.001).In addition,the GR nuclear translocation,and the protein expression level of Hsp90 that assisted the nuclear translocation of GR were increased after CORT treatment.Conclusion Glucocorticoids inhibit the apoptosis induced by serum deprivation in mouse H22 hepatocarcinoma cells,and the mechanism is associated with the enhanced GR nuclear translocation assisted by Hsp90.

关 键 词：血清剥夺 糖皮质激素受体 细胞凋亡 HSP90 

分 类 号：R735.7[医药卫生—肿瘤]