Related Experiment Video
Updated: May 20, 2026

Co-immunoprecipitation Assay for Studying Functional Interactions Between Receptors and Enzymes
Published on: September 28, 2018
PKCδ regulates death receptor 5 expression induced by PS-341 through ATF4-ATF3/CHOP axis in human lung cancer cells
Linyan Xu1, Ling Su, Xiangguo Liu
1Shandong University School of Life Sciences, Room 103, South Building, 27 Shanda South Road, Jinan, Shandong 250100, China.
Abstract:
PS-341 (bortezomib), a proteasome inhibitor, has been approved for the treatment of multiple myeloma. Our previous work has shown that PS-341 induces death receptor 5 (DR5)-dependent apoptosis and enhances the TNF-related apoptosis-inducing ligand-induced apoptosis in human non-small cell lung cancer cells. However, the definite mechanism remains undefined. In the present study, we reveal that PKCδ and RSK2 mediate PS-341-induced DR5 upregulation, involving coactivation of endoplasmic reticulum (ER) stress. We discovered that PS-341 activated ER stress through elevating the expression of BiP, p-eIF2α, IRE1α, ATF4, ATF3, and CCAAT/enhancer-binding protein homologous protein (CHOP). Further study showed that DR5 upregulation was dependent on ATF4, ATF3, and CHOP expression. Silencing either one of the ATF4, ATF3, and CHOP expression decreased DR5 upregulation and subsequent apoptosis. We determined that ATF4 regulated ATF3 and CHOP expression. Thereafter, ATF3 and CHOP formed a complex and regulated DR5 expression. In addition, we discovered that the phosphorylation of extracellular signal-regulated kinase (ERK) 1/2 and RSK2 were elevated after PS-341 treatment and inhibition of their phosphorylation using MAP-ERK kinase 1/2 inhibitor decreased the DR5 level, indicating that ERK/RSK2 signaling is involved in DR5 upregulation. Furthermore, we detected the cleavage of PKCδ, and the blockage of PKCδ expression cut down DR5 upregulation and apoptosis. Importantly, knockdown of PKCδ expression decreased the induction of ER stress and the phosphorylation of ERK1/2 and RSK2, suggesting that PKCδ regulates DR5 expression through ERK/RSK2 signaling and ATF4-CHOP/ATF3 axis. Collectively, we show that PS-341 induces PKCδ-dependent DR5 expression through activation of ERK/RSK2 and ER stress signaling pathway.
Insights
Bortezomib (PS-341) triggers cancer cell death by upregulating death receptor 5 (DR5) via PKCδ, ERK/RSK2, and ER stress pathways, involving ATF4, ATF3, and CHOP. This clarifies the mechanism of bortezomib-induced apoptosis.
Area of Science:
- Oncology
- Molecular Biology
- Cellular Signaling
Background:
- Bortezomib (PS-341) is an approved proteasome inhibitor for multiple myeloma.
- Previous studies indicated PS-341 induces apoptosis in non-small cell lung cancer (NSCLC) cells via death receptor 5 (DR5).
- The precise molecular mechanisms underlying PS-341-induced DR5 upregulation and apoptosis in NSCLC remain incompletely understood.
Purpose of the Study:
- To elucidate the signaling pathways and molecular mediators involved in PS-341-induced DR5 upregulation and apoptosis in NSCLC.
- To investigate the roles of protein kinase C delta (PKCδ), ribosomal S6 kinase 2 (RSK2), and endoplasmic reticulum (ER) stress in this process.
Main Methods:
- Cellular assays including gene silencing (siRNA) and Western blotting to assess protein expression and phosphorylation.
- Inhibition of specific signaling pathways using kinase inhibitors (e.g., MAP-ERK kinase 1/2 inhibitor).
- Analysis of ER stress markers (BiP, p-eIF2α, IRE1α, ATF4, ATF3, CHOP) and DR5 expression levels following PS-341 treatment.
Main Results:
- PS-341 treatment activated ER stress, evidenced by increased BiP, p-eIF2α, IRE1α, ATF4, ATF3, and CHOP expression.
- DR5 upregulation was dependent on ATF4, ATF3, and CHOP, with ATF4 regulating ATF3/CHOP, and ATF3/CHOP forming a complex to control DR5.
- PKCδ cleavage and activation of ERK/RSK2 signaling were crucial for PS-341-induced DR5 upregulation and apoptosis, with PKCδ regulating ER stress and ERK/RSK2 phosphorylation.
Conclusions:
- PS-341 induces cancer cell apoptosis by upregulating DR5 expression.
- This upregulation is mediated by a novel pathway involving PKCδ, ERK/RSK2 signaling, and ER stress activation.
- The ATF4-ATF3/CHOP axis plays a critical role in translating ER stress signals to DR5 expression, elucidating a key mechanism of bortezomib's anti-cancer effects.
Related Concept Videos
The Extrinsic Apoptotic Pathway
The Intrinsic Apoptotic Pathway
PI3K/mTOR/AKT Signaling Pathway
MAPK Signaling Cascades
NF-κB-dependent Signaling Pathway
NF-κB-dependent Signaling Mechanism
The heterodimer of NF-κB...
Overview of Cell Death
Cell death was observed in the early 19th century, but there was no experimental evidence to prove it. In 1842, Carl Vogt first discovered cell death in a metamorphic toad; however, it was not termed ‘cell death.’ Scientists discovered different cell death pathways only in the 20th century...
