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目的分析抗肺癌单克隆抗体16D3靶抗原蛋白在人肺癌细胞及组织中的表达,鉴定16D3对肺癌的体内外抑制功能,为肺癌靶向治疗提供候选抗体药物。方法采用免疫组化分析16D3靶蛋白在人肺癌组织中的表达;采用细胞免疫荧光检测单克隆抗体16D3识别的抗原在肺癌细胞中的表达及亚细胞定位;共接种移植瘤模型评估16D3对肺癌生长的抑制作用,Transwell法和CCK-8法检测单抗16D3对人肺癌细胞迁移、侵袭和增殖能力的影响;采用流式细胞术检测16D3靶蛋白分别在亲本和sphere培养的人肺癌细胞中的表达。结果单克隆抗体16D3的靶蛋白在7种人肺癌细胞的胞内及胞膜上均有表达,在76.9%人肺癌患者组织中特异上调表达。该单抗在体内能较显著地抑制人肺腺癌GLC-82的生长,抑制率为58.23%(P<0.05)。在体外对该细胞的增殖、迁移和侵袭具有一定的抑制作用,抑制率分别为26.91%、24.23%和23.67%。该单抗的靶蛋白膜表达阳性细胞比例经sphere培养后显著上升。结论单抗16D3靶抗原在人肺癌中特异表达,并可被sphere培养富集。单抗16D3在体外具有多种抑制功能,在体内能显著抑制人肺腺癌生长,可能是一个肺癌靶向治疗的潜在新药。
Objective To analyze the expression of anti-lung cancer monoclonal antibody 16D3 target protein in human lung cancer cells and tissues, and to identify the anti-tumor effect of 16D3 against lung cancer in vitro and in vivo, so as to provide candidate antibody drugs for lung cancer targeted therapy. Methods The expression of 16D3 target protein in human lung cancer tissues was analyzed by immunohistochemistry. The expression and subcellular localization of 16D3 antigen in lung cancer cells were detected by immunofluorescence staining. The inoculated xenografts model was used to evaluate the effect of 16D3 on the growth of lung cancer The effects of 16D3 monoclonal antibody on the migration, invasion and proliferation of human lung cancer cells were detected by Transwell assay and CCK-8 assay. The expression of 16D3 target protein in human lung cancer cells cultured in vitro and in vivo were detected by flow cytometry . Results The target protein of monoclonal antibody 16D3 was expressed both intracellular and intracellularly in 7 kinds of human lung cancer cells, and was specifically up-regulated in 76.9% human lung cancer tissues. The monoclonal antibody could significantly inhibit the growth of human lung adenocarcinoma GLC-82 in vivo with the inhibition rate of 58.23% (P <0.05). In vitro proliferation, migration and invasion of the cells have a certain inhibitory effect, the inhibition rates were 26.91%, 24.23% and 23.67%. The proportion of positive cells expressing the target protein membrane of the monoclonal antibody increased significantly after the sphere culture. Conclusion McAb 16D3 target antigen is specifically expressed in human lung cancer and can be enriched by sphere culture. McAb 16D3 has a variety of inhibitory functions in vitro, can significantly inhibit the growth of human lung adenocarcinoma in vivo, may be a potential new drug targeted therapy of lung cancer.