摘要
目的:对实验室已构建的新型免疫毒素IAB-TAT-BH3在肝癌靶向治疗中的作用进行初步评价。方法:首先通过免疫组化技术对人肝癌组织芯片中CA125的表达进行检测,同时利用RT-PCR、Western blot和流式细胞术从RNA和蛋白水平对肝癌细胞中CA125的表达进行系统分析;然后通过Western blot和流式细胞术(Flow cytometry,FCM)检测免疫毒素IABTAT-BH3与肝癌细胞株SMMC-7721的结合能力,最后采用MTT法检测其对于SMMC-7721的靶向杀伤作用。结果:CA125在人类肝癌组织芯片和肝癌细胞株SMMC-7721中呈现高表达;免疫毒素IAB-TAT-BH3对于SMMC-7721具有良好的结合能力,且与两种对照免疫毒素相比,具有更强的细胞杀伤作用,IC50值约0.3μmol/L。结论:膜蛋白CA125是潜在的肝癌分子靶标,靶向CA125的新型免疫毒素IAB-TAT-BH3在体外对肝癌细胞具有一定的特异性杀伤作用,在肝癌靶向治疗上具有潜在的应用前景。
Objective:To appraise the effect of novel immunotoxin IAB-TAT-BH3 which was constructed by our lab in hepato- carcinoma targeted therapy. Methods: First, a human liver carcinoma tissue array was used to investigate the expression of CA125 by immunohistochemistry. Then expression of CA125 in different hepatocarcinoma cells was measured by RT-PCR, Western blot and Flow cytometry. Finally, the binding of the immunotoxin to liver cancer cell SMMC-7721 was detected by Western blot and Flow cytometry and the cytotoxicity of 1AB-TAT-BH3 on SMMC-7721 was further appraised by MTT. Results: CA125 was highly expressed in most human liver carcinoma tissues and SMMC-7721 cells; the immunotoxin IAB-TAT-BH3 could specifically binding to SMMC-7721 cells and the IC50 of IAB-TAT-BH3 to this cell line was about 0.3 p, mol/L which is much better than the other two control immunotoxins. Conclusion: The membrane protein CA125 has potential to be a novel marker of hepatocarcinoma. The novel immunotoxin lAB-TAT- BH3 which can target to CA125 can specifically inhibit the proliferation of liver cancer cells in vitro, has great potential in hepatocawi- noma targeted therapy.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2014年第3期342-346,共5页
Chinese Journal of Immunology
基金
上海市科委生物医药领域重点科技攻关专项(074319104)资助
关键词
CA125
分子靶标
肝癌
靶向治疗
CA125
Molecular target
Hepatocarcinoma
Targeted therapy