Abstract
The activation of receptor-interacting protein kinase 1 (RIPK1) by death-inducing signaling complex (DISC) formation is essential for triggering the necroptotic mode of cell death under apoptosis-deficient conditions. Thus, targeting the induction of necroptosis by modulating RIPK1 activity could be an effective strategy to bypass apoptosis resistance in certain types of cancer. In this study, we screened a series of arborinane triterpenoids purified from Rubia philippinesis and identified rubiarbonol B (Ru–B) as a potent caspase-8 activator that induces DISC-mediated apoptosis in multiple types of cancer cells. However, in RIPK3-expressing human colorectal cancer (CRC) cells, the pharmacological or genetic inhibition of caspase-8 shifted the mode of cell death by Ru–B from apoptosis to necroptosis though upregulation of RIPK1 phosphorylation. Conversely, Ru–B-induced cell death was almost completely abrogated by RIPK1 deficiency. The enhanced RIPK1 phosphorylation and necroptosis triggered by Ru–B treatment occurred independently of tumor necrosis factor receptor signaling and was mediated by the production of reactive oxygen species via NADPH oxidase 1 in CRC cells. Thus, we propose Ru–B as a novel anticancer agent that activates RIPK1-dependent cell death via ROS production, and suggest its potential as a novel necroptosis-targeting compound in apoptosis-resistant CRC.
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The data sets used or analyzed during the current study are available from the corresponding author on reasonable request.
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Abbreviations
- BH:
-
Butylated hydroxyanisole
- CCCP:
-
Carbonyl cyanide m-chlorophenyl hydrazine
- CHX:
-
Cycloheximide
- cIAP1/2:
-
Cellular inhibitor of apoptosis 1 and 2
- DHE:
-
Dihydroethidium
- DISC:
-
Death-inducing signaling complex
- DPI:
-
Diphenyleneiodonium
- FADD:
-
Fas-associated death domain
- IKK:
-
IkB kinase
- LUBAC:
-
Linear ubiquitin chain assembly complex
- MLKL:
-
Mixed lineage kinase-domain-like
- NADPH:
-
Nicotinamide adenine dinucleotide phosphate
- Nec-1:
-
Necrostain-1
- NF-kB:
-
Nuclear factor kappa B
- NOX:
-
NADPH oxidase
- NSA:
-
Necrosulfonamide
- O2 − :
-
Superoxide
- PCD:
-
Programmed cell death
- PI:
-
Propidium iodide
- RDA:
-
RIPK1-dependent apoptosis
- RIPK1:
-
Receptor-interacting protein kinase 1
- ROS:
-
Reactive oxygen species
- Ru-B:
-
Rubiarbonol B
- SM:
-
SM-164 (smac mimetics)
- TNF:
-
Tumor necrosis factor
- TRADD:
-
TNFR1-associated death domain protein
- z-DEVD:
-
Z-DEVD-fmk (benzyloxy-carbonyl-Asp-(OMe)-Glu-(OMe)-Val-Asp-(OMe)-fluoromethyl ketone)
- z-IETD:
-
Z-IETD-fmk (benzyloxy-carbonyl-Ile-Glu-(OMe)-Tyr-Asp-(OMe)-fluoromethyl ketone)
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Acknowledgements
We thank Dr. A. Justin Rucker (Duke University, USA) for assistance in the preparation of this manuscript
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This work as supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIP) (No. 2020R1A2C2005317; No. 2017R1A5A2015385; No. 2019R1I1A1A01061239, 2020R1A2B5B01002415).
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HSB participated in the design of the study, carried out bench experiments, and analyzed data. Both EJ and KAP contributed to the final version of manuscript. K-C S and CSJ helped carrying out bench experiments related to this study. JHH, HR, and HYL helped drafting the manuscript by providing critical intellectual input. KTQ, IP, and MN carried out the experiments for the purification of Ru–B and NMR spectra data analysis. MN and GMH designed this study and wrote the manuscript with comments from the coauthors, and all authors collaborated on the work.
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Byun, H.S., Ju, E., Park, K.A. et al. Rubiarbonol B induces RIPK1-dependent necroptosis via NOX1-derived ROS production. Cell Biol Toxicol 39, 1677–1696 (2023). https://doi.org/10.1007/s10565-022-09774-6
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DOI: https://doi.org/10.1007/s10565-022-09774-6