Effect of heat shock protein 90α on Cyclin B1 in L02 cells under oxidative stress
DUAN Dan-ping1, HUANG Ting-yuan2, LIU Chen-yang3, LI Yan3, LI Yan-ru1, ZOU Fei3, WANG Zhi1, CHEN Xue-mei3
1. Guangzhou Hospital for Occupational Disease Prevention and Treatment, Key Laboratory of Occupational Environment and Health, Guangzhou Twelfth People’s Hospital, Guangzhou, Guangdong 510620, China; 2. Department of Chronic Non-communicable Disease Prevention and Control, Guangzhou Municipal Center for Disease Control and Prevention, Guangzhou, Guangdong 510440, China; 3. Department of Occupational Health and Occupational Medicine, School of Public Health, Southern Medical University, Guangzhou, Guangdong 510515, China
Abstract:Objective To observe the changes of heat shock protein 90α (Hsp90α) and Cyclin B1 in L02 cells under the condition of oxidative stress, to detect the changes of CyclinB1 activity after down-regulated expression of Hsp90α, to identify the effect of Hsp90α on the function of CyclinB1, and to provide a novel idea for prevention and control of liver disease related to oxidative stress. Methods We established a model for oxidative stress in L02 cells induced by 200 μM H2O2, and then used CCK-8 method to observe the effect of oxidative stress on cell survival. The intracellular levels of glutathione (GSH) were observed by colorimetric method. Flow cytometer was used to examine the cell cycle. The effect of oxidative stress on intracellular synthesis of Hsp90α and CyclinB1 was detected by Western-blotting. Results Under 200 μM H2O2 concentration, the intracellular levels of GSH were negatively time dependent, and the antioxidant capacity was continuously reduced. Compared with the control group, the intracellular levels of GSH significantly decreased after 6 hours stimulation (all P<0.05). The inhibition of cell proliferation was time dependent. G2/M phase arrest in L02 cells after oxidative stress was significant. Oxidative stress led to first increased and then decreased intracellular expression of Hsp90α. The expression of Hsp90α decreased significantly after 24 hours stimulation, while Cyclin B1 expression increased significantly, showing statistically significant differences as compared with the control group (both P<0.05). Conclusion Oxidative stress can dynamically affect the intracellular expression of Hsp90α and Cyclin B1,suggesting that in this process, Hsp90 plays a protective role, reduces the direct damage caused by stress, and maintains the stability of Cyclin B1 function.
段丹萍, 黄婷苑, 刘晨阳, 李艳, 李燕茹, 邹飞, 王致, 陈雪梅. 人肝细胞氧化应激模型中热休克蛋白90α对细胞周期蛋白B1的降解调控机制研究[J]. 实用预防医学, 2023, 30(2): 152-155.
DUAN Dan-ping, HUANG Ting-yuan, LIU Chen-yang, LI Yan, LI Yan-ru, ZOU Fei, WANG Zhi, CHEN Xue-mei. Effect of heat shock protein 90α on Cyclin B1 in L02 cells under oxidative stress. , 2023, 30(2): 152-155.
[1] Pan X, Wen SW, Kaminga AC, et al.Gut metabolites and inflammation factors in non-alcoholic fatty liver disease: a systematic review and meta-analysis[J].Sci Rep,2020,10(1):8848. [2] Tu W, Wang H, Li S, et al.The anti-inflammatory and anti-oxidant mechanisms of the Keap1/Nrf2/ARE signaling pathway in chronic diseases[J].Aging Dis,2019,10(3):637-651. [3] 赵杰,齐永芬,鱼艳荣.氧化应激在肝纤维化发生发展中的作用[J].临床肝胆病杂志,2019,35(9):2067-2071. [4] Jia P, Ji S, Zhang H, et al.Piceatannol ameliorates hepatic oxidative damage and mitochondrial dysfunction of weaned piglets challenged with diquat[J].Animals (Basel),2020,10(7):1239. [5] 郑坤,嘎鲁,马宇衡,等.活性氧(ROS)依赖性抗肿瘤药物的研究进展[J].广东药科大学学报,2022,38(1):130-136. [6] 刘巍,徐兆发,郭美欣,等.莱菔硫烷对汞致大鼠肝脏氧化应激的拮抗作用研究[J].实用预防医学,2016,23(6):671-674. [7] 窦博,马重阳,侯伟欣,等.基于TLR4/NF-κB/iNOS通路的截断逆挽方药物血清对L02细胞氧化应激损伤的影响[J].中国中医药信息杂志,2022,29(4):87-91. [8] Chebotareva N, Bobkova I, Shilov E.Heat shock proteins and kidney disease: perspectives of HSP therapy[J].Cell Stress Chaperones,2017,22(3):319-343.[9] Birbo B, Madu EE, Madu CO, et al.Role of HSP90 in cancer[J].Int J Mol Sci,2021,22(19):10317. [10] Ikwegbue PC, Masamba P, Oyinloye BE, et al.Roles of heat shock proteins in apoptosis, oxidative stress, human inflammatory diseases, and cancer[J].Pharmaceuticals (Basel),2017,11(1):2. [11] 林晶晶,张倍宁,姜楠,等.热休克蛋白在创伤愈合过程中的作用研究进展[J].中华卫生应急电子杂志,2017,3(6):375-377. [12] 马庆荣,余佩芝,张帆,等.热疗中热休克蛋白90对26S蛋白酶体的调控机制[J].南方医科大学学报,2017,37(4):537-541. [13] 刘谦,路名芝,刘勇.细胞周期标记物在肿瘤中的应用[J].临床与实验病理学杂志,2012,28(2):192-196. [14] 李武杰,李勇强,张樊苹,等.Claudin-1与cyclinB1在下咽鳞状细胞癌中的表达及其临床意义[J].中国耳鼻咽喉颅底外科杂志,2020,26(5):537-543. [15] 林万尊.靶向阻断cyclinB1对鼻咽癌细胞自噬的影响及其分子机制研究[D].福州:福建医科大学,2019. [16] 陈雪梅,戴沛娟,段丹萍,等.人肝癌细胞氧化应激模型中热休克蛋白90α对20S蛋白酶体功能的影响[J].临床医学工程,2012,19(7):1056-1058. [17] Zuo L, Zhou T, Pannell BK, et al.Biological and physiological role of reactive oxygen species--the good, the bad and the ugly[J].Acta Physiol (Oxf),2015,214(3):329-348. [18] Sadasivam N, Kim YJ, Radhakrishnan K, et al.Oxidative stress, genomic integrity, and liver diseases[J].Molecules,2022,27(10):3159. [19] Padmini E, Usha Rani M.Heat-shock protein 90 alpha (HSP90α) modulates signaling pathways towards tolerance of oxidative stress and enhanced survival of hepatocytes of Mugil cephalus[J].Cell Stress Chaperones,2011,16(4):411-425. [20] Sarvani C, Sireesh D, Ramkumar KM.Unraveling the role of ER stress inhibitors in the context of metabolic diseases[J].Pharmacol Res,2017,119:412-421. [21] Oroz J,Kim JH,Chang BJ,et al.Mechanistic basis for the recognition of a misfolded protein by the molecular chaperone Hsp90[J].Nat Struct Mol Biol,2017,24(4):407-413.