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Bortezomib overcomes TRAIL resistance in Burkitt's lymphoma by enhancing apoptosis via reactive oxygen species‑mediated DR5 upregulation and MAPK pathway activation

  • Authors:
    • Jiajie Chen
    • Yujia You
    • Wei Zhao
    • Yichuan Zhou
    • Shiqi Pu
    • Xiaoyu Bai
    • Ping Yang
    • Minhui Li
  • View Affiliations / Copyright

    Affiliations: School of Basic Medicine, Chengdu Medical College, Chengdu, Sichuan 610500, P.R. China, School of Laboratory Medicine, Chengdu Medical College, Chengdu, Sichuan 610500, P.R. China, School of Bioscience and Technology, Chengdu Medical College, Chengdu, Sichuan 610500, P.R. China
    Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 588
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    Published online on: October 14, 2025
       https://doi.org/10.3892/ol.2025.15334
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Abstract

Tumor necrosis factor‑related apoptosis‑inducing ligand (TRAIL) selectively kills tumor cells and exerts minimal toxic effects on normal cells. However, resistance to TRAIL‑induced apoptosis is a major obstacle in the clinical application of TRAIL. Bortezomib can enhance the tumor‑killing effect of TRAIL in certain tumors, but understanding in Burkitt's lymphoma (BL), especially in TRAIL‑resistant BL cells, is limited. Therefore, the present study aimed to assess the synergistic effect of bortezomib and TRAIL in BL. Using the Raji cell line, the least susceptible to TRAIL among BL cell lines, the present study assessed the effect and mechanism by which bortezomib reverses resistance to TRAIL. Cell proliferation inhibition was assessed using the Cell Counting Kit‑8 assay. Apoptosis was assessed by flow cytometry. Intracellular reactive oxygen species (ROS) were detected with DCFH‑DA, and mitochondrial membrane potential was measured using JC‑1. Expression levels of apoptosis proteins and MAPK signaling pathway‑related proteins were analyzed by western blot. Combination of bortezomib and TRAIL strongly and synergistically inhibited Raji and CA46 BL cell proliferation. Furthermore, the combination of bortezomib and TRAIL was associated with the following: Induction of apoptosis; increased levels of ROS; presence of mitochondrial membrane potential disorders; upregulation of the levels of apoptosis‑related proteins cleaved caspase 8/9/3 and cleaved poly (ADP‑ribose) polymerase; and downregulation of the levels of antiapoptotic factors Bcl‑2 and Bcl‑xl. In addition, bortezomib induced a significant increase in the expression levels of death receptor 5 (DR5), a TRAIL receptor. Pretreatment with the antioxidant N‑acetylcysteine inhibited not only ROS upregulation but also DR5 upregulation induced by bortezomib in Raji cells. Furthermore, the present study revealed that the combination of bortezomib and TRAIL could regulate the levels of MAPK signaling pathway‑related proteins, such as phosphorylated extracellular signal‑regulated kinase 1/2, phosphorylated (p‑)p38, p‑c‑Jun, p‑activating transcription factor 2 and phosphorylated stress‑activated protein kinase/c‑Jun N‑terminal kinase. Therefore, the results indicate that bortezomib may enhance Raji cell sensitivity to TRAIL via ROS‑dependent upregulation of DR5, induce apoptosis through the MAPK signaling pathway, and subsequently inhibit cell proliferation. Additionally, bortezomib combined with TRAIL had a potential synergistic apoptosis‑inducing effect in TRAIL‑resistant BL cells.
View Figures

Figure 1

Bortezomib combined with TRAIL
synergistically inhibits the proliferation of Burkitt's lymphoma
cells. (A) Raji and CA46 cells were incubated with different
concentrations of TRAIL for 24 h, and cell proliferation was
assessed using a CCK-8 assay. (B) Raji and (C) CA46 cells were
incubated with different concentrations of bortezomib alone or in
combination with 100 ng/ml TRAIL for 24 h, and cell proliferation
was evaluated using a CCK-8 assay. TRAIL, tumor necrosis
factor-related apoptosis-inducing ligand; CCK-8, Cell Counting
Kit-8; Bort, bortezomib.

Figure 2

Morphological characterization of
Raji cells after combination treatment with bortezomib and TRAIL.
The combination of TRAIL and bortezomib was compared with TRAIL or
bortezomib alone (indicated by arrows; magnification, ×10). R1
indicates a high magnification image (×40) showing apoptotic
features, including cell shrinkage, membrane rupture and apoptotic
body formation. Scale bars, 20 and 5 µm (R1). TRAIL, tumor necrosis
factor-related apoptosis-inducing ligand; Bort, bortezomib.

Figure 3

Bortezomib combined with TRAIL
induces apoptosis in Raji cells. (A) Apoptotic effects of
bortezomib and/or TRAIL in Raji cells were determined using flow
cytometry via dual staining with annexin V-FITC and PI. (B)
Expression levels of apoptosis-related proteins were assessed using
western blot. TRAIL, tumor necrosis factor-related
apoptosis-inducing ligand; Bort, bortezomib.

Figure 4

Bortezomib combined with TRAIL
induces apoptosis via the mitochondrial pathway in Raji cells. (A)
Changes of MMP in Raji cells were detected using flow cytometry.
(B) Expression levels of mitochondrial pathway-related proteins
were detected using western blot analysis. TRAIL, tumor necrosis
factor-related apoptosis-inducing ligand; Bort, bortezomib; MMP,
mitochondrial membrane potential.

Figure 5

Bortezomib combined with TRAIL
induces ROS production in Raji cells. (A) Intracellular ROS levels
were assessed using the fluorescence dye DCFH-DA and flow cytometry
analysis. (B) Changes in ROS were evaluated using histogram
analysis. TRAIL, tumor necrosis factor-related apoptosis-inducing
ligand; Bort, bortezomib; ROS, reactive oxygen species; NAC,
N-acetylcysteine.

Figure 6

Effect of bortezomib combined with
TRAIL on DR5 via ROS in Raji cells. (A) Expression of DR5 on the
cell surface was detected using flow cytometry analysis with
PE-CD262 (DR5) monoclonal antibodies. (B) Western blot was used to
detect CHOP protein expression levels. TRAIL, tumor necrosis
factor-related apoptosis-inducing ligand; Bort, bortezomib; DR5,
death receptor 5; ROS, reactive oxygen species; CHOP, C/EBP
homologous protein; NAC, N-acetylcysteine.

Figure 7

Effect of bortezomib combined with
TRAIL on the MAPK signaling pathway in Raji cells. (A) MAPK
signaling pathway relative protein expression in Raji cells treated
with bortezomib and/or TRAIL was detected using western blot
analysis. (B) Histogram analysis of protein expression. TRAIL,
tumor necrosis factor-related apoptosis-inducing ligand; Bort,
bortezomib; SAPK/JNK, stress-activated protein kinase/c-Jun
N-terminal kinase; ATF2, activating transcription factor 2; ERK,
extracellular signal-regulated kinase; p-, phosphorylated.
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Spandidos Publications style
Chen J, You Y, Zhao W, Zhou Y, Pu S, Bai X, Yang P and Li M: Bortezomib overcomes TRAIL resistance in Burkitt's lymphoma by enhancing apoptosis via reactive oxygen species‑mediated DR5 upregulation and MAPK pathway activation. Oncol Lett 30: 588, 2025.
APA
Chen, J., You, Y., Zhao, W., Zhou, Y., Pu, S., Bai, X. ... Li, M. (2025). Bortezomib overcomes TRAIL resistance in Burkitt's lymphoma by enhancing apoptosis via reactive oxygen species‑mediated DR5 upregulation and MAPK pathway activation. Oncology Letters, 30, 588. https://doi.org/10.3892/ol.2025.15334
MLA
Chen, J., You, Y., Zhao, W., Zhou, Y., Pu, S., Bai, X., Yang, P., Li, M."Bortezomib overcomes TRAIL resistance in Burkitt's lymphoma by enhancing apoptosis via reactive oxygen species‑mediated DR5 upregulation and MAPK pathway activation". Oncology Letters 30.6 (2025): 588.
Chicago
Chen, J., You, Y., Zhao, W., Zhou, Y., Pu, S., Bai, X., Yang, P., Li, M."Bortezomib overcomes TRAIL resistance in Burkitt's lymphoma by enhancing apoptosis via reactive oxygen species‑mediated DR5 upregulation and MAPK pathway activation". Oncology Letters 30, no. 6 (2025): 588. https://doi.org/10.3892/ol.2025.15334
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Spandidos Publications style
Chen J, You Y, Zhao W, Zhou Y, Pu S, Bai X, Yang P and Li M: Bortezomib overcomes TRAIL resistance in Burkitt's lymphoma by enhancing apoptosis via reactive oxygen species‑mediated DR5 upregulation and MAPK pathway activation. Oncol Lett 30: 588, 2025.
APA
Chen, J., You, Y., Zhao, W., Zhou, Y., Pu, S., Bai, X. ... Li, M. (2025). Bortezomib overcomes TRAIL resistance in Burkitt's lymphoma by enhancing apoptosis via reactive oxygen species‑mediated DR5 upregulation and MAPK pathway activation. Oncology Letters, 30, 588. https://doi.org/10.3892/ol.2025.15334
MLA
Chen, J., You, Y., Zhao, W., Zhou, Y., Pu, S., Bai, X., Yang, P., Li, M."Bortezomib overcomes TRAIL resistance in Burkitt's lymphoma by enhancing apoptosis via reactive oxygen species‑mediated DR5 upregulation and MAPK pathway activation". Oncology Letters 30.6 (2025): 588.
Chicago
Chen, J., You, Y., Zhao, W., Zhou, Y., Pu, S., Bai, X., Yang, P., Li, M."Bortezomib overcomes TRAIL resistance in Burkitt's lymphoma by enhancing apoptosis via reactive oxygen species‑mediated DR5 upregulation and MAPK pathway activation". Oncology Letters 30, no. 6 (2025): 588. https://doi.org/10.3892/ol.2025.15334
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