title: Knockdown of miR-182 promotes apoptosis via regulating RIP1 deubiquitination in TNF-α-treated triple-negative breast cancer cells.
abstract: Overexpression of microRNA-182 (miR-182) is found in multiple cancers, but the association of miR-182 expression with the sensitivity of triple-negative breast cancer (TNBC) cells to tumor necrosis factor-alpha (TNF-α) remains unknown. In this study, up-regulation of miR-182 was validated in TNBC patients and cell lines. Knockdown of miR-182 was observed to hinder the proliferation of BT-549 cells. More importantly, knockdown of miR-182 significantly promoted the apoptosis induced by TNF-α treatment in BT-549. JC-1 staining and western blot assays revealed that the K63-linked ubiquitin chains on receptor-interacting protein 1 (RIP1) were removed and the outer mitochondrial membrane potential (MMP) and permeability was altered upon combination of TNF-α with anti-miR-182. We then demonstrated that knockdown of miR-182 up-regulated the expression of cylindromatosis (CYLD) deubiquitinase, which promoted the formation of death-inducing signaling complex (DISC) and subsequent caspase-8 activation in TNF-α-treated BT-549 cells. Collectively, the results of the present study improve our understanding of the role of miR-182 in TNBC, knockdown of which facilitates the degradation of ubiquitin chains on RIP1, leading to the caspase-8 activation and apoptosis in TNF-α-treated TNBC cells. This may be valuable for the development of cancer therapy.
authors.affiliation: Department of Breast Surgery, First Affiliated Hospital of Zhejiang Chinese Medical University, Youdian Road 54#, Hangzhou, China.
authors.forename: Like
authors.initials: L
authors.lastname: Wo
authors.affiliation: College of Life Science, Zhejiang Chinese Medical University, Hangzhou, 310053, China.
authors.forename: Dezhao
authors.initials: D
authors.lastname: Lu
authors.affiliation: Department of Breast Surgery, First Affiliated Hospital of Zhejiang Chinese Medical University, Youdian Road 54#, Hangzhou, China. hzxidonggu@163.com.
authors.forename: Xidong
authors.initials: X
authors.lastname: Gu
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D049109:Cell Proliferation / Q000187:drug effects; D005260:Female; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007124:Immunoenzyme Techniques; D047468:Immunoprecipitation; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000235:genetics*; D009367:Neoplasm Staging; D028861:Nuclear Pore Complex Proteins / Q000235:genetics / Q000378:metabolism*; D011379:Prognosis; D012333:RNA, Messenger / Q000235:genetics; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D064726:Triple Negative Breast Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D025801:Ubiquitin / Q000378:metabolism*; D054875:Ubiquitination / Q000235:genetics*
chemical_list: C119724:AGFG1 protein, human; D014408:Biomarkers, Tumor; D035683:MicroRNAs; C538984:Mirn182 microRNA, human; D028861:Nuclear Pore Complex Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D014409:Tumor Necrosis Factor-alpha; D025801:Ubiquitin
keywords: CYLD; Deubiquitination; TNBC; TNF-α; miR-182