Key necroptotic proteins are required for Smac mimetic-mediated sensitization of cholangiocarcinoma cells to TNF-α and chemotherapeutic gemcitabine-induced necroptosis.

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  • Additional Information
    • Source:
      Publisher: Public Library of Science Country of Publication: United States NLM ID: 101285081 Publication Model: eCollection Cited Medium: Internet ISSN: 1932-6203 (Electronic) Linking ISSN: 19326203 NLM ISO Abbreviation: PLoS One Subsets: MEDLINE
    • Publication Information:
      Original Publication: San Francisco, CA : Public Library of Science
    • Subject Terms:
      Bridged Bicyclo Compounds, Heterocyclic/*pharmacology ; Deoxycytidine/*analogs & derivatives ; Necroptosis/*drug effects ; Receptor-Interacting Protein Serine-Threonine Kinases/*metabolism ; Triazoles/*pharmacology ; Tumor Necrosis Factor-alpha/*pharmacology; Biliary Tract Neoplasms/metabolism ; Biliary Tract Neoplasms/pathology ; Caspase Inhibitors/pharmacology ; Cell Cycle Checkpoints/drug effects ; Cell Line, Tumor ; Cholangiocarcinoma/metabolism ; Cholangiocarcinoma/pathology ; Deoxycytidine/pharmacology ; Drug Synergism ; Humans ; Protein Kinases/chemistry ; Protein Kinases/genetics ; Protein Kinases/metabolism ; RNA Interference ; RNA, Small Interfering/metabolism ; Receptor-Interacting Protein Serine-Threonine Kinases/genetics ; Tumor Necrosis Factor-alpha/genetics ; Tumor Necrosis Factor-alpha/metabolism ; Gemcitabine
    • Abstract:
      Cholangiocarcinoma (CCA), a malignant tumor originating in the biliary tract, is well known to be associated with adverse clinical outcomes and high mortality rates due to the lack of effective therapy. Evasion of apoptosis is considered a key contributor to therapeutic success and chemotherapy resistance in CCA, highlighting the need for novel therapeutic strategies. In this study, we demonstrated that the induction of necroptosis, a novel regulated form of necrosis, could potentially serve as a novel therapeutic approach for CCA patients. The RNA sequencing data in The Cancer Genome Atlas (TCGA) database were analyzed and revealed that both receptor-interacting protein kinase 3 (RIPK3) and mixed lineage kinase domain-like (MLKL), two essential mediators of necroptosis, were upregulated in CCA tissues when compared with the levels in normal bile ducts. We demonstrated in a panel of CCA cell lines that RIPK3 was differentially expressed in CCA cell lines, while MLKL was more highly expressed in CCA cell lines than in nontumor cholangiocytes. We therefore showed that treatment with both tumor necrosis factor-α (TNF-α) and Smac mimetic, an inhibitor of apoptosis protein (IAP) antagonist, induced RIPK1/RIPK3/MLKL-dependent necroptosis in CCA cells when caspases were blocked. The necroptotic induction in a panel of CCA cells was correlated with RIPK3 expression. Intriguingly, we demonstrated that Smac mimetic sensitized CCA cells to a low dose of standard chemotherapy, gemcitabine, and induced necroptosis in an RIPK1/RIPK3/MLKL-dependent manner upon caspase inhibition but not in nontumor cholangiocytes. We further demonstrated that Smac mimetic and gemcitabine synergistically induced an increase in TNF-α mRNA levels and that Smac mimetic reversed gemcitabine-induced cell cycle arrest, leading to cell killing. Collectively, our present study demonstrated that TNF-α and gemcitabine induced RIPK1/RIPK3/MLKL-dependent necroptosis upon IAP depletion and caspase inhibition; therefore, our findings have pivotal implications for designing a novel necroptosis-based therapeutic strategy for CCA patients.
      Competing Interests: The authors have declared that no competing interests exist.
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    • Grant Information:
      MR/N01247X/1 United Kingdom MRC_ Medical Research Council
    • Accession Number:
      0 (Bridged Bicyclo Compounds, Heterocyclic)
      0 (Caspase Inhibitors)
      0 (RNA, Small Interfering)
      0 (SM 164)
      0 (Triazoles)
      0 (Tumor Necrosis Factor-alpha)
      0W860991D6 (Deoxycytidine)
      EC 2.7.- (MLKL protein, human)
      EC 2.7.- (Protein Kinases)
      EC 2.7.11.1 (RIPK1 protein, human)
      EC 2.7.11.1 (RIPK3 protein, human)
      EC 2.7.11.1 (Receptor-Interacting Protein Serine-Threonine Kinases)
      0 (Gemcitabine)
    • Publication Date:
      Date Created: 20200109 Date Completed: 20200413 Latest Revision: 20221207
    • Publication Date:
      20240829
    • Accession Number:
      PMC6948742
    • Accession Number:
      10.1371/journal.pone.0227454
    • Accession Number:
      31914150