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

作　　者：袁翔[1] 姜琳琳[1] 曹善楠[1] 张孝云[1] 齐怀丰 熊冬生[1] 廖晓龙[1] 

机构地区：[1]中国医学科学院北京协和医学院血液学研究所实验血液学国家重点实验室,天津300020 [2]天津红日金博达生物技术有限公司

出　　处：《白血病．淋巴瘤》2013年第8期462-465,共4页Journal of Leukemia & Lymphoma

基　　金：国家自然科学基金（30971291）

摘　　要：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.

关 键 词：增强融合蛋白 抗CD20抗体 BJAB细胞 细胞周期阻滞 

分 类 号：R733.1[医药卫生—肿瘤]