A cell-based study on the efficacy of lenalidomide combined with zanubrutinib against central nervous system lymphoma

doi:10.19803/j.1672-8629.20260208

Abstract

Abstract: Objective To explore the combined effects of lenalidomide and zanubrutinib on the viability, apoptosis, proliferation, and cycle of OCI-Ly10 cells. Methods The CCK-8 method was used to observe the inhibitory effects of 50 μmol·L^-1 lenalidomide, 0.312 5 μmol·L^-1 zanubrutinib (monotherapy group), 50 μmol·L^-1 lenalidomide in combination with 0.312 5 μmol·L^-1 zanubrutinib (combination group), and 20、40、80、160、320 μmol·L^-1 methotrexate (MTX) on OCI-Ly10 cells. Annexin V-FITC&PI staining was used to detect the effects on the apoptosis of OCI-Ly10 cells in the monotherapy groups and the combination group. 5-ethynyl-2′-deoxyuridine (EdU) staining was adopted to observe the effects on the proliferation of OCI-Ly10 cells in these groups. PI staining was employed to evaluate the effects on the cycle of OCI-Ly10 cells in these groups. Results Lenalidomide (50 μmol·L^-1) and zanubrutinib (0.312 5 μmol·L^-1) exerted an inhibitory effect on cell viability 24 hours after being administered alone. In contrast, cell viability was significantly suppressed in the combination group (both P<0.01). Notably, no statistically significant difference in cell viability was observed between the combination group and 80 μmol·L^-1 MTX. Compared with the control group, apoptosis in OCI-Ly10 cells was significantly aggravated in monotherapy groups (both P<0.001). Moreover, the pro-apoptotic effect was more pronounced in the combination group than in monotherapy groups (both P<0.001). Lenalidomide had no significant effect on OCI-Ly10 cells compared with the control group while zanubrutinib significantly inhibited cell proliferation (P<0.001). The inhibitory effect on cell proliferation was stronger in the combination group than in monotherapy groups (both P<0.001). Cell cycle analysis found that monotherapy significantly increased the proportion of cells in the G₀/G₁phase (both P<0.001) but decreased the proportion in the S phase (both P<0.001) compared with the control group. Compared with lenalidomide monotherapy, the combination treatment elevated the proportion of cells in the G₀/G₁ phase (P<0.01) and reduced the proportion in the S phase (P<0.001). Conversely, compared with zanubrutinib monotherapy, the combination treatment significantly decreased the proportion of cells in the G₀/G₁ phase (P<0.001) but increased the proportion in the S phase (P<0.01). Conclusion After 24 hours of drug intervention, lenalidomide alone can promote cell apoptosis without affecting cell viability or cell proliferation while zanubrutinib alone can not only induce cell apoptosis but also inhibit cell proliferation by arresting cells in the G₀/G₁ phase. The combination of both drugs can exert a synergistic effect by significantly reducing cell viability and producing an inhibitory effect comparable to 80 μmol·L^-1 MTX so that cell apoptosis is enhanced and cell proliferation is inhibited.

Key words: Lenalidomide, Zanubrutinib, Primary Central Nervous System Lymphoma(PCNSL), Bruton Tyrosine Kinase (BTK), OCI-Ly10 Cells, Apoptosis, Cycle, Proliferation

CLC Number:

Liu Dan, Gao Lei, Zhang Min, Ren Xiaohui, Liu Qian. A cell-based study on the efficacy of lenalidomide combined with zanubrutinib against central nervous system lymphoma[J]. Chinese Journal of Pharmacovigilance, 2026, 23(10): 1086-1091.

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