| 范冬梅,杨圆圆,姜琳琳.融合蛋白anti-CD19(Fab)-LDM对B细胞淋巴瘤BJAB细胞杀伤作用及其机制研究[J].中国肿瘤,2019,28(3):220-226. |
| 融合蛋白anti-CD19(Fab)-LDM对B细胞淋巴瘤BJAB细胞杀伤作用及其机制研究 |
| Cytotoxicity of Engineered Fusion Protein Anti-CD19(Fab)-LDM on B-cell Lymphoma BJAB Cells and Its Mechanism |
| 投稿时间：2018-09-05 |
| DOI：10.11735/j.issn.1004-0242.2019.03.A011 |
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| 中文关键词: 基因工程抗体 融合蛋白 力达霉素 B细胞淋巴瘤 细胞周期 |
| 英文关键词:genetic engineered antibody fusion protein lidamycin B-cell lymphoma cell cycle |
| 基金项目:国家科技重大专项（2012ZX09102301） |
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| 中文摘要: |
| 摘 要：［目的］ 探讨融合蛋白anti-CD19(Fab)-LDM对B细胞淋巴瘤细胞株BJAB细胞的杀伤作用及其机制研究。［方法］ 利用流式细胞分析技术（FACS）和激光共聚焦显微镜技术，检测融合蛋白anti-CD19(Fab)-LDM与BJAB细胞的结合活性。MTT法检测融合蛋白anti-CD19(Fab)-LDM对BJAB细胞的体外杀伤活性。彗星电泳实验检测融合蛋白anti-CD19(Fab)-LDM对BJAB细胞DNA的损伤。FACS检测不同浓度融合蛋白anti-CD19(Fab)-LDM处理BJAB细胞后细胞周期的变化。 ［结果］ FACS和激光共聚焦显微镜技术实验结果表明，融合蛋白anti-CD19(Fab)-LDM能与BJAB细胞结合。MTT法实验结果表明，融合蛋白anti-CD19(Fab)-LDM对BJAB细胞杀伤活性较单用力达霉素或阿霉素强，IC50值分别为(0.15±0.02)nmol/L、(0.38±0.03)nmol/L 和(57.15±2.30)nmol/L。彗星电泳实验结果表明，用5pmol/L的融合蛋白anti-CD19(Fab)-LDM及力达霉素处理BJAB细胞后，可引起细胞不同程度的DNA损伤，由于融合蛋白anti-CD19(Fab)-LDM具有靶向性和细胞内化的特性，对DNA造成的损伤较力达霉素组明显。FACS结果显示，随着融合蛋白anti-CD19(Fab)-LDM浓度的增加，S期的比例从53.78%升高至77.29%，呈剂量依赖性。［结论］ 融合蛋白anti-CD19(Fab)-LDM可以靶向杀伤B细胞淋巴瘤细胞株BJAB，引起细胞周期阻滞，在B细胞淋巴瘤生物治疗中具有潜在的应用价值。 |
| 英文摘要: |
| Abstract：［Purpose］ To investigate the cytotoxicity of the engineered fusion protein anti-CD19 (Fab)-LDM to B-cell lymphoma BJAB cells and its mechanism.［Methods］ The antigen binding activity of anti-CD19(Fab)-LDM on BJAB cells was observed by FACS and confocal microscopy. The cytotoxicity of the engineered fusion protein anti-CD19(Fab)-LDM to BJAB cells was tested by MTT assay. Comet assay was used to determine the DNA damage in BJAB cells induced by anti-CD19(Fab)-LDM. Changes of cell cycle of BJAB cells treated with anti-CD19(Fab)-LDM were detected by FACS. ［Results］ FACS and confocal microscopy showed that the antibody bound specifically to BJAB cells. MTT assay demonstrated that the engineered fusion protein anti-CD19(Fab)-LDM strongly enhanced the cytotoxicity to BJAB cells compared with adriamycin or lidamycin (LDM). The comet assay showed that cells treated with anti-CD19(Fab)-LDM induced more DNA damage than cells treated with LDM. BJAB cells treated with anti-CD19(Fab)-LDM resulted in cell cycle arrest in a concentration dependence manner.［Conclusion］ The fusion protein anti-CD19(Fab)-LDM can target to kill CD19-positive B lymphoma cells and induce cell cycle arrest. These findings may be useful in clinical practice for B-cell lymphoma treatment. |
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