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[Bortezomib Inhibits Extranodal Natural Killer/T Cell Lymphoma, Nasal Type by Targeting NF-κB Signaling Pathway]
Jian-Hua Li1, Li Zhang2, You Feng3
1Department of Oncology, West China Hospital, Sichuan University, Chengdu 610041, China.
Objective:
To investigate the anti-tumor effect of bortezomib on extranodal natural killer/T cell lymphoma, nasal type (ENKTL).
Methods:
SNK-6 cells were treated with different mass concentrations of bortezomib (0, 1, 2, 4, 5, 6 ng/mL) for 24, 48, 72 h, and different concentrations of nuclear factor-kappa B (NF-κB) signaling pathway inhibitor BAY11-7082 (0, 1, 2, 2.5, 5, 10, 20 μmol/L) for 24 h respectively, then the cell viability was measured by CCK8 kit and the half inhibitory concentration (IC 50) was calculated. SNK-6 cells were treated with 30μmol/L Z-VAD-FMK (Pan-caspase inhibitor)+3ng/mL bortezomib, and 5, 10 μmol/L BAY11-7082+3 ng/mL bortezomib for 24 h respectively, then the cell viability was measured by CCK8 kit. After treatment of SNK-6 cells with different mass concentrations of bortezomib for 24 h, apoptosis was detected by AnnexinⅤ/PI flow cytometry; the expression of apoptosis-related protein Caspase-3, poly ADP-ribose polymerase (PARP) and Bcl-2 and NF-κB signaling pathway key proteins P65 and P100/P52 were detected by Western blot.
Results:
Bortezomib inhibited the proliferation of SNK-6 cells in a dose-dependent manner ( P<0.05), and IC 50( (2.87±0.06) ng/mL) at 24 h was lower than that at 48 h and 72 h ( P<0.05). BAY11-7082 also inhibited the proliferation of SNK-6 cells with an IC 50= (9.73±0.36) μmol/L at 24 h. The combination treatment indicated that Z-VAD-FMK could attenuate the inhibitory effect of bortezomib on the proliferation of SNK-6 cells ( P<0.05), while BAY11-7082 could enhance the inhibitory effect of bortezomib on the proliferation of SNK-6 cells ( P<0.05). After treatment of SNK-6 cells with bortezomib for 24 h, apoptosis-related protein Caspase-3 cleavage, PARP activation, and Bcl-2 cleavage; NF-κB signaling pathway-related protein P65 phosphorylation level decreased, and P52 decreased.
Conclusion:
Bortezomib inhibits ENKTL cells proliferation by inhibiting NF-κB signaling pathway and induces apoptosis of ENKTL cells via mitochondria-mediated caspase pathway.
Insights
Bortezomib effectively inhibits extranodal natural killer/T cell lymphoma (ENKTL) cell proliferation and induces apoptosis. This occurs through the inhibition of the nuclear factor-kappa B (NF-κB) signaling pathway and activation of the caspase pathway.
Area of Science:
- Oncology
- Immunology
- Molecular Biology
Background:
- Extranodal natural killer/T cell lymphoma, nasal type (ENKTL) is an aggressive non-Hodgkin lymphoma.
- Targeted therapies are crucial for improving outcomes in ENKTL.
- The role of the nuclear factor-kappa B (NF-κB) signaling pathway in ENKTL pathogenesis is increasingly recognized.
Purpose of the Study:
- To evaluate the anti-tumor effects of bortezomib on ENKTL cells.
- To investigate the underlying mechanisms, including apoptosis induction and NF-κB pathway modulation.
Main Methods:
- SNK-6 ENKTL cells were treated with varying concentrations of bortezomib and BAY11-7082 (NF-κB inhibitor).
- Cell viability was assessed using CCK8 assays to determine IC50 values.
- Apoptosis was analyzed via Annexin V/PI staining and Western blot for key proteins (Caspase-3, PARP, Bcl-2, P65, P52).
Main Results:
- Bortezomib demonstrated dose-dependent inhibition of SNK-6 cell proliferation with a low IC50 (2.87 ng/mL at 24h).
- BAY11-7082 also inhibited cell proliferation (IC50 = 9.73 μmol/L at 24h).
- Bortezomib induced apoptosis, evidenced by Caspase-3 and PARP activation, and decreased Bcl-2 levels. It also reduced NF-κB pathway activity (decreased P65 phosphorylation and P52 levels).
Conclusions:
- Bortezomib exhibits significant anti-tumor activity against ENKTL cells.
- The drug inhibits ENKTL cell proliferation by suppressing the NF-κB signaling pathway.
- Bortezomib induces apoptosis in ENKTL cells through a mitochondria-mediated caspase-dependent pathway.
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