A non-metabolic function of hexokinase 2 in small cell lung cancer:promotes cancer cell stemness by increasing USP11-mediated CD133 stability  

作　　者：Juhong Wang Fei Shao Yannan Yang Wei Wang Xueying Yang Renda Li Hong Cheng Sijin Sun Xiaoli Feng Yibo Gao Jie He Zhimin Lu 

机构地区：[1]Department of Thoracic Surgery,National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100021,P.R.China [2]State Key Laboratory of Molecular Oncology,National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100021,P.R.China [3]Laboratory of Translational Medicine,National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100021,P.R.China [4]Department of Medical Oncology,National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100021,P.R.China [5]Department of Pathology,National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Beijing 100021,P.R.China [6]Central Laboratory,National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital&Shenzhen Hospital,Chinese Academy of Medical Sciences and Peking Union Medical College,Shenzhen,Guangdong 518116,P.R.China [7]Zhejiang Provincial Key Laboratory of Pancreatic Disease,The First Affiliated Hospital and Institute of Translational Medicine,Zhejiang University School of Medicine,Hangzhou,Zhejiang 310029,P.R.China [8]Cancer Center,Zhejiang University,Hangzhou,Zhejiang 310029,P.R.China

出　　处：《Cancer Communications》2022年第10期1008-1027,共20页癌症通讯（英文）

基　　金：Ministry of Science and Technology of the People’s Republic of China,Grant/Award Number:2020YFA0803300;National Natural Science Foundation of China,Grant/Award Numbers:82188102,82030074,82122053,32100574;Beijing Municipal Science&Technology Commission,Grant/Award Number:Z191100006619115;R&D Program of Beijing Municipal Education commission,Grant/Award Number:KJZD20191002302;CAMS Innovation Fund for Medical Science,Grant/Award Numbers:2021-1-I2M-012,2021-I2M-1-067;Non-profit Central Research Institute Fund of Chinese Academy of Medical Sciences,Grant/Award Number:2021-PT310-001;Key-Area Research and Development Program of Guangdong Province,Grant/Award Number:2021B0101420005;Sanming Project of Medicine in Shenzhen,Grant/Award Numbers:SZSM201612097,SZSM201812062;Aiyou Foundation,Grant/Award Number:KY201701;Natural Science Foundation of Shandong Province,Grant/Award Number:ZR2020QH191;Zhejiang Natural Science Foundation-Key Project,Grant/Award Number:LD21H160003;Leading Innovative and Entrepreneur Team Introduction Program of Zhejiang,Grant/Award Number:2019R01001;Zhimin Lu is the Kuancheng Wang Distinguished Chair。

摘　　要：Background:Maintenance of cancer stem-like cell(CSC)stemness supported by aberrantly regulated cancer cell metabolism is critical for CSC self-renewal and tumor progression.As a key glycolytic enzyme,hexokinase 2(HK2)plays an instrumental role in aerobic glycolysis and tumor progression.However,whether HK2 directly contribute to CSC stemness maintenance in small cell lung cancer(SCLC)is largely unclear.In this study,we aimed to investgate whether HK2 independent of its glycolytic activity is directly involved in stemness maintenance of CSC in SCLC.Methods:Immunoblotting analyses were conducted to determine the expression of HK2 in SCLC CSCs and their differentiated counterparts.CSC-like properties and tumorigenesis of SCLC cells with or without HK2 depletion or overexpression were examined by sphere formation assay and xenograft mouse model.Immunoprecipitation and mass spectrometry analyses were performed to identify the binding proteins of CD133.The expression levels of CD133-associated and CSC-relevant proteins were evaluated by immunoblotting,immunoprecipitation,immunofluorescence,and immunohistochemistry assay.RNA expression levels of Nanog,POU5F1,Lin28,HK2,Prominin-1 were analyzed through quantitative reverse transcription PCR.Polyubiquitination of CD133 was examined by in vitro or in vivo ubiquitination assay.CD133+cells were sorted by flow cytometry using an anti-CD133 antibody.Results:We demonstrated that HK2 expression was much higher in CSCs of SCLC than in their differentiated counterparts.HK2 depletion inhibited CSC stemness and promoted CSC differentiation.Mechanistically,nonmitochondrial HK2 directly interacted with CD133 and enhanced CD133 expression without affecting CD133 mRNA levels.The interaction of HK2 and CD133 promoted the binding of the deubiquitinase ubiquitin-specific protease 11(USP11)to CD133,thereby inhibiting CD133 polyubiquitylation and degradation.HK2-mediated upregulation of CD133 expression enhanced the expression of cell renewal regulators,SCLC cell stemness,and tumor growth in

关 键 词：HK2 metabolic enzyme non-metabolic function cancer stem-like cell CD133 USP11 UBIQUITYLATION SCLC 

分 类 号：R734.2[医药卫生—肿瘤]