强化融合蛋白抗CD20(Fab)-力达霉素对淋巴瘤BJAB细胞增殖和DNA损伤的作用

摘要

Objective To investigate the growth inhibition and DNA damage of energized fusion protein anti-CD20(Fab)-LDM on B JAB cells in vitro.Methods The binding activity of fusion protein anti-CD20 (Fab)-LDP to B JAB cells was studied by flow cytometry and confocal laser scanning microscopy.MTT assay was used to study the energized fusion protein anti-CD20(Fab)-LDM on cell growth of B JAB cells.Comet assay was employed to detect DNA damage in B JAB cells.The cell growth cycle of BJAB was analyzed by FACS.Results The recombinant fusion protein anti-CD20 (Fab)-LDP possessed an significant target affinity towarded BJAB cells.The energized fusion protein anti-CD20(Fab)-LDM showed obvious inhibition on proliferation,as well as induced potent DNA damage in B JAB cells in vitro compared with lidamycin.B JAB cells treated with energized fusion protein anti-CD20 (Fab)-LDM showed S phase cell cycle.Conclusion The energized fusion protein anti-CD20 (Fab)-LDM could target binding to BJAB cells and significantly inhibit the proliferation of B JAB cells by inducing DNA damage and S phase arrest.%目的 研究强化融合蛋白抗CD20(Fab)-力达霉素(LDM)体外对CD20表达阳性的淋巴瘤细胞株B JAB增殖和DNA损伤的作用.方法 流式细胞术检测融合蛋白抗CD20(Fab)-力达霉素辅基蛋白(LDP)与BJAB细胞的结合活性；激光共聚焦显微镜观察融合蛋白抗CD20(Fab)-LDP靶向结合B JAB细胞情况；MTF法检测强化融合蛋白抗CD20(Fab)-LDM对BJAB细胞的生长抑制作用；单细胞凝胶电泳(SCGE)检测强化融合蛋白抗CD20(Fab)-LDM对DNA的损伤；流式技术检测强化融合蛋白抗CD20(Fab)-LDM诱导BJAB细胞的细胞周期的改变.结果 融合蛋白抗CD20(Fab)-LDP与BJAB细胞结合活性为阳性；激光共聚焦显微镜下观察,融合蛋白抗CD20(Fab)-LDP靶向结合在BJAB细胞表面；强化融合蛋白抗CD20(Fab)-LDM作用于淋巴瘤BJAB细胞后,可以显著抑制BJAB细胞的生长,引起DNA损伤,诱导S期阻滞.结论 强化融合蛋白抗CD20(Fab)-LDM可以靶向结合淋巴瘤BJAB细胞,并且具有显著的生长抑制作用,其作用机制是引起DNA的损伤.