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目的:鉴定单克隆抗体MS57-2.1(简称MS57-2.1单抗)的胃癌特异性,研究MS57-2.1单抗的体外和体内抑瘤功能,为胃癌的靶向治疗提供候选抗体药物。方法:本课题组前期利用4个胃癌细胞株膜蛋白免疫A/J小鼠,通过杂交瘤联合高通量流式细胞技术筛选并获得抗胃癌MS57-2.1单抗。通过流式细胞和ELISA检测MS57-2.1单抗的胃癌特异性。通过免疫荧光技术鉴定MS57-2.1单抗的靶抗原在胃癌细胞的定位。通过体外和体内实验初步研究MS57-2.1单抗抑制胃癌细胞移动以及侵袭的作用。结果:流式细胞检测结果表明MS57-2.1单抗可以高亲和力与特定胃癌细胞株结合,而几乎不与正常人外周血细胞结合。ELISA结果表明MS57-2.1单抗可与胃癌组织膜蛋白结合,而不与幽门螺杆菌及胃癌细胞分泌蛋白结合。免疫荧光检测表明MS57-2.1单抗的靶抗原定位于胃癌细胞膜表面。在Transwell小室迁移和侵袭实验中,MS57-2.1单抗处理后胃癌细胞MKN45和BGC823移动能力受到明显抑制,穿过小室细胞数和穿膜细胞数显著低于无关同型单抗对照组和空白对照组。在裸鼠实验中,MS57-2.1单抗处理组的裸鼠肿瘤播散程度显著低于无关同型单抗对照组和空白对照组。结论:MS57-2.1单抗具有抑制胃癌细胞迁移、侵袭和播散的作用,是胃癌靶向治疗具有潜在应用价值的候选抗体药物。
OBJECTIVE: To identify the gastric cancer specificity of monoclonal antibody MS57-2.1 (MS57-2.1 monoclonal antibody for short), to study the anti-tumor function of MS57-2.1 monoclonal antibody in vitro and in vivo and to provide candidate antibody drugs for the targeted therapy of gastric cancer. Methods: A group of A / J mice were immunized with membrane proteins of four gastric cancer cell lines in our group and screened by hybridoma combined with high-throughput flow cytometry to obtain anti-gastric cancer MS57-2.1 monoclonal antibody. Gastric Cancer Specificity of MS57-2.1 Monoclonal Antibodies Detected by Flow Cytometry and ELISA. The localization of the target antigen of the MS57-2.1 mAb in gastric cancer cells was identified by immunofluorescence. In vitro and in vivo experiments preliminary study of MS57-2.1 monoclonal antibody inhibition of gastric cancer cell migration and invasion. Results: The results of flow cytometry showed that MS57-2.1 mAb could bind to specific gastric cancer cell lines with high affinity and hardly bind with normal human peripheral blood cells. ELISA results show that MS57-2.1 monoclonal antibody can bind to the membrane protein of gastric cancer, but not with Helicobacter pylori and gastric cancer cell secreted protein binding. Immunofluorescence showed that the target antigen of MS57-2.1 mAb localized on the surface of gastric cancer cell membrane. In Transwell chamber migration and invasion experiments, the migration ability of gastric cancer cells MKN45 and BGC823 was significantly inhibited after MS57-2.1 monoclonal antibody treatment, the number of cells crossing the cell and the number of transmembrane cells was significantly lower than that of the non-related iso-mAb control group and the blank control group . In the nude mice experiment, the degree of tumor dissemination in nude mice treated with MS57-2.1 mAb was significantly lower than that in the non-related homo-mAb control group and the blank control group. Conclusion: MS57-2.1 mAb can inhibit the migration, invasion and dissemination of gastric cancer cells and is a potential antibody candidate for targeted therapy of gastric cancer.