功能性单克隆抗体40E11靶向治疗肺癌的实验研究

Targeting therapy of lung cancer using functional monoclonal antibody 40E11

目的:检测分析抗肺癌单克隆抗体40E11识别的靶抗原在肺癌细胞系及肺癌组织中的表达,研究单克隆抗体40E11体内外抑瘤功能,并初步鉴定其识别的抗原蛋白,为肺癌的靶向治疗提供候选抗体药物及靶标。方法:应用细胞免疫荧光检测单克隆抗体40E11所识别的靶抗原在肺癌细胞系中的表达及定位,免疫组化检测其在人肺癌组织中的表达;CCK-8法检测分析40E11对肺癌细胞增殖的影响。裸鼠体内治疗实验研究40E11对移植瘤生长的抑制作用。Western bloting鉴定该单克隆抗体识别的抗原的分子量。结果:单克隆抗体40E11识别的抗原在人肺腺癌细胞(A549、GLC-82、ANIP-973)、人肺鳞癌细胞(GLC-P)的胞内及胞膜均有表达;该抗原在84.6%的人肺癌组织中的表达较癌旁组织显著上调;CCK-8法检测结果显示单克隆抗体40E11明显抑制肺癌细胞体外生长;抗体治疗实验结果表明单克隆抗体40E11明显抑制体内肺癌细胞生长,其抑制率达64.4%,P<0.05;蛋白质印迹法显示40E11识别的抗原相对分子质量约为100×103。结论:抗肺癌单克隆抗体40E11在体内外能显著抑制肺癌细胞的生长,为肺癌的靶向治疗提供具有重要应用价值的候选抗体药物。

OBJECTIVE： To study the expression of the targeted antigen of anti-lung cancer monoclonal antibody 40E11 in lung cancer, and study the inhibitory function of 40E11 mAb against lung cancer in vivo, and in vitro and identify the antigen, so as to provide candidate target and antibody for cancer cell-targeted therapy of lung cancer. METHODS： The expression and localization of targeted antigen of 40E11 mAb in lung cancer cell lines and lung cancer tissues were examined by immunofluorescence assay and immunohistochemistry. The inhibitory function of 40E11 mAb against lung cancer cell lines in vivo and in vitro was studied by tumor treatment experiment and CCK-8 assay. The corresponding antigen of 40E11 was identified by western bloting. RESULTS： The targeted antigen of 40E11 mAb was expressed in the cytoplasm and cell surface of 4 lung cancer lines, including human lung adenocarcinoma cell lines （A549, GLC-82, ANIP 973） and squamous carcinoma cell lines（GLC-P）. The expression of the targeted antigen was significantly upregu- lated in about 84.6% of human lung cancer tissues. The 40E11 mAb could obviously suppress proliferation of lung cancer cells in vivo and in vitro, with the inhibitory rate being 64. 4 M （P〈0.05）. Western bloting revealed that the molecular weight of the antigen of 40E11 antibody was about 100×10^3. CONCLUSION： 40E11 mAb can significantly inhibit growth of lung cancer in vivo and in vitro, and 40E11 mAb could be a candidate drug for the targeted therapy of lung cancer.