Mechanisms underlying reversed TRAIL sensitivity in acquired bortezomib-resistant non-small cell lung cancer cells  

作　　者：Leonie De Wilt Bartosz Kamil Sobocki Gerrit Jansen Hessan Tabeian Steven de Jong Godefridus J.Peters Frank Kruyt 

机构地区：[1]Department of Medical Oncology,Amsterdam University Medical Centers,Location VUMC,Vrije Universiteit Amsterdam,Amsterdam 1007MB,the Netherlands [2]Department of Biochemistry,Medical University of Gdańsk,Gdańsk 80-210,Poland [3]Department of Rheumatology,Amsterdam University Medical Centers,Vrije Universiteit Amsterdam,Amsterdam 1081 HV,the Netherlands [4]Department of Medical Oncology,University of Groningen,University Medical Center Groningen,Groningen 9713 GZ,the Netherlands

出　　处：《Cancer Drug Resistance》2024年第1期869-889,共21页癌症耐药(英文)

基　　金：grant VU2006-3567 from the Dutch Cancer Society.

摘　　要：Aim:The therapeutic targeting of the tumor necrosis factor(TNF)-related apoptosis-inducing ligand(TRAIL)death receptors in cancer,including non-small cell lung cancer(NSCLC),is a widely studied approach for tumor selective apoptotic cell death therapy.However,apoptosis resistance is often encountered.The main aim of this study was to investigate the apoptotic mechanism underlying TRAIL sensitivity in three bortezomib(BTZ)-resistant NSCLC variants,combining induction of both the intrinsic and extrinsic pathways.Methods:Sensitivity to TRAIL in BTZ-resistant variants was determined using a tetrazolium(MTT)and a clonogenic assay.A RT-qPCR profiling mRNA array was used to determine apoptosis pathway-specific gene expression.The expression of these proteins was determined through ELISA assays and western Blotting,while apoptosis(sub-G1)and cytokine expression were determined using flow cytometry.Apoptotic genes were silenced by specific siRNAs.Lipid rafts were isolated with fractional ultracentrifugation.Results:A549BTZR(BTZ-resistant)cells were sensitive to TRAIL in contrast to parental A549 cells,which are resistant to TRAIL.TRAIL-sensitive H460 cells remained equally sensitive for TRAIL as H460BTZR.In A549BTZR cells,we identified an increased mRNA expression of TNFRSF11B[osteoprotegerin(OPG)]and caspase-1,-4 and-5 mRNAs involved in cytokine activation and immunogenic cell death.Although the OPG,interleukin-6(IL-6),and interleukin-8(IL-8)protein levels were markedly enhanced(122-,103-,and 11-fold,respectively)in the A549BTZR cells,this was not sufficient to trigger TRAIL-induced apoptosis in the parental A549 cells.Regarding the extrinsic apoptotic pathway,the A549BTZR cells showed TRAIL-R1-dependent TRAIL sensitivity.The shift of TRAIL-R1 from non-lipid into lipid rafts enhanced TRAIL-induced apoptosis.In the intrinsic apoptotic pathway,a strong increase in the mRNA and protein levels of the anti-apoptotic myeloid leukemia cell differentiation protein(Mcl-1)and B-cell leukemia/lymphoma 2(Bcl-2)was found,whereas the

关 键 词：TRAIL BORTEZOMIB resistance SENSITIZATION Bcl-2 family lipid rafts CYTOKINES 

分 类 号：R73[医药卫生—肿瘤]