来那度胺联合泽布替尼抗中枢神经系统淋巴瘤的细胞研究

doi:10.19803/j.1672-8629.20260208

摘要/Abstract

摘要： 目的探讨来那度胺和泽布替尼两药联合对人弥漫大B细胞淋巴瘤细胞系OCI-Ly10细胞活性、凋亡、增殖、周期影响。方法利用CCK-8法观察50 μmol·L^-1来那度胺,0.312 5 μmol·L^-1泽布替尼（单药组）,50 μmol·L^-1来那度胺联合0.312 5 μmol·L^-1泽布替尼（联合组）,20、40、80、160、320 μmol·L^-1甲氨蝶呤（Methotrexate, MTX）对OCI-Ly10细胞活性抑制情况;采用Annexin V-FITC&PI双染检测单药组及联合组对OCI-Ly10细胞凋亡影响;采用5-乙炔基-2′-脱氧尿苷（EdU）染色检测单药组及联合组对OCI-Ly10细胞增殖影响;采用PI染色检测单药组及联合组对OCI-Ly10细胞周期影响。结果 50 μmol·L^-1来那度胺、0.312 5 μmol·L^-1泽布替尼干预OCI-Ly10 24 h对细胞活性均无抑制作用,联合组活性受到显著抑制（P均<0.01）;联合组与80 μmol·L^-1 MTX相比,差异无统计学意义。与正常组相比,单药组显著促进OCI-Ly10细胞凋亡（P均<0.001）;与单药组相比,联合组显著促进细胞凋亡（P均<0.001）。与正常组相比,来那度胺对OCI-Ly10细胞增殖无影响,泽布替尼显著抑制细胞增殖（P<0.001）;与单药组相比,联合组细胞增殖显著受到抑制（P均<0.001）。与正常组相比,单药组G₀/G₁期显著升高（P均<0.001）、S期显著降低（P均<0.001）;与来那度胺相比,联合组G₀/G₁期显著升高（P<0.01）,S期显著降低（P<0.001）;与泽布替尼相比,联合组G₀/G₁期显著降低（P<0.001）,S期显著升高（P<0.01）。结论经药物干预24 h,来那度胺单药可促进细胞凋亡,但对细胞活力及增殖无显著影响;泽布替尼单药不仅能诱导细胞凋亡,还可通过将细胞阻滞于G₀/G₁期抑制其增殖。两药联合呈现协同效应,显著降低细胞活力,其抑制效果与80 μmol·L^-1MTX相当,进一步增强凋亡诱导作用并抑制细胞增殖。

关键词: 来那度胺, 泽布替尼, 原发性中枢神经系统淋巴瘤, 布鲁顿酪氨酸激酶, OCI-Ly10细胞, 凋亡, 周期, 增殖

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

中图分类号:

刘丹, 高磊, 张敏, 任晓辉, 刘潜. 来那度胺联合泽布替尼抗中枢神经系统淋巴瘤的细胞研究[J]. 中国药物警戒, 2026, 23(10): 1086-1091.

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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