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Absence of death receptor translocation into lipid rafts in acquired TRAIL-resistant NSCLC cells
Wen Ouyang1, Chunxu Yang, Simin Zhang
1Department of Radiation and Medical Oncology, Zhongnan Hospital, Wuhan University, Wuhan 430071, P.R. China.
Abstract:
Resistance to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a major limitation for its clinical use. The mechanisms of TRAIL resistance have been mostly studied in the context of cell lines that are intrinsically resistant to TRAIL. However, little is known about the molecular alterations that contribute to the development of acquired resistance during treatment with TRAIL. In this study, we established H460R, an isogenic cell line with acquired TRAIL resistance, from the TRAIL‑sensitive human lung cancer cell line H460 to investigate the mechanisms of acquired resistance. The acquired TRAIL‑resistant H460R cells remained sensitive to cisplatin. The mRNA and protein expression levels of death receptor 4 (DR4) and death receptor 5 (DR5) were not altered in either of the TRAIL-treated cell lines. Nevertheless, tests in which the DR4 or DR5 gene was overexpressed or silenced suggest that death receptor expression is necessary but not sufficient for TRAIL‑induced apoptosis. Compared with parental TRAIL-sensitive H460 cells, H460R cells showed a decreased TRAIL-induced translocation of DR4/DR5 into lipid rafts. Further studies showed that nystatin partially prevented lipid raft aggregation and DR4 and DR5 clustering and reduced apoptosis in H460 cells again. Analysis of apoptotic molecules showed that more pro-caspase-8, FADD, caspase-3 and Bid, but less cFLIP in H460 cells than in H460R cells. Our findings suggest that the lack of death receptor redistribution negatively impacts DISC assembly in lipid rafts, which at least partially leads to the development of acquired resistance to TRAIL in H460R cells.
Insights
Acquired resistance to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) involves impaired death receptor translocation into lipid rafts. This hinders death-inducing signaling complex assembly, contributing to TRAIL resistance in cancer cells.
Area of Science:
- Cancer Biology
- Molecular Oncology
- Apoptosis Research
Background:
- Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) therapy is limited by drug resistance.
- Mechanisms of acquired TRAIL resistance are poorly understood, unlike intrinsic resistance.
- Investigating acquired resistance is crucial for improving TRAIL-based cancer treatments.
Purpose of the Study:
- To elucidate the molecular mechanisms underlying acquired resistance to TRAIL.
- To establish and characterize an isogenic cell line model of acquired TRAIL resistance.
- To identify key molecular alterations contributing to TRAIL unresponsiveness.
Main Methods:
- Established H460R, a TRAIL-resistant cell line, from TRAIL-sensitive H460 lung cancer cells.
- Assessed TRAIL-induced apoptosis, death receptor (DR4/DR5) expression, and translocation.
- Utilized gene overexpression/silencing and nystatin treatment to investigate lipid raft involvement.
- Analyzed expression of key apoptotic pathway molecules (caspases, FADD, Bid, cFLIP).
Main Results:
- H460R cells acquired TRAIL resistance but remained cisplatin-sensitive.
- DR4/DR5 expression levels were unchanged, but their TRAIL-induced translocation to lipid rafts decreased in H460R cells.
- Nystatin treatment impaired lipid raft aggregation and reduced apoptosis in sensitive cells.
- H460R cells exhibited altered levels of apoptotic proteins, including less cFLIP compared to parental cells.
Conclusions:
- Acquired TRAIL resistance is associated with impaired death receptor redistribution into lipid rafts.
- Reduced death receptor translocation negatively impacts DISC assembly, contributing to TRAIL resistance.
- Understanding these mechanisms may offer new therapeutic strategies to overcome TRAIL resistance in cancer.
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