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

Effect of energized fusion protein anti-CD20 （Fab）-LDM on proliferation and DNA damage of human lymphoma cell line

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BJAB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BJAB细胞情况；M_rr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。

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-CD：o （Fab）-LDP to B JAB cells was studied by flow cytometry and confocal laser scanning microscopy. MT3 assay was used to study the energized fusion protein anti-CDao（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 B JAB 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-CDao（Fab）-LDM showed obvious inhibition on proliferation, as well as induced potent DNA damage in BJAB cells in vitro compared with lidamyein. B JAB cells treated with energized fusion protein anti-CD20 （Fab）-LDM showed S phase cell cycle. Conclusion The energized fusion protein auti-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.