目的： 制备抑制食管癌细胞与肺血管内皮黏附的功能性单克隆抗体，为研究食管癌转移的靶向治疗剂打下基础。方法： 以人食管癌新鲜组织分离的肿瘤细胞免疫BALB/c小鼠，将免疫小鼠脾细胞与SP2/0细胞融合后，采用活细胞荧光、ELISA、免疫组化、肿瘤细胞 内皮细胞黏附实验、Weastern bloting等方法筛选并鉴定抑制肿瘤细胞黏附的功能性单克隆抗体。结果： 共融合获得了1 134株杂交瘤克隆，在能与食管癌细胞膜反应的486株克隆中，有294株与正常食管上皮不反应或低反应；筛选到15株单抗显著地抑制食管癌细胞与人正常肺血管内皮黏附，其中2株单抗识别的黏附分子相对分子质量分别为30 000和60 000。结论 ： 成功获得了多株具有抑制食管癌细胞与肺血管内皮黏附的功能性抗体，其中2株单抗具有治疗食管癌转移的潜在应用价值。

Objective: To prepare monoclonal antibody for inhibition of adhesion between esophageal carcinoma and pulmonary vascular endothelia, so as to pave a way for the targeted treatment of esophageal carcinoma metastasis.Methods: Tumor cells were separated from esophageal carcinoma tissues and were used to immunize Balb/c mouse. The spleen cells of the immunized mouse were fused with SP2/0 cells. The anti-adhesion antibodies were then screened and identified by ELISA, immunohistochemistry, immunofluorescence, tumor-endothelial cell adhesion assay, and Western blotting. Results: Totally 1 134 clones of monoclonal antibodies were obtained after fusion. Among the 486 clones which could interact with the membrane of the esophageal carcinoma, 294 clones had no or weak reactions with normal esophageal epithelia. The 15 selected clones significantly suppressed the adherence between esophageal cancer cells and lung endothelia. Western blotting displayed that 2 of the clones reacted with the antigens having a molecular weight of 30 000 and 60 000, respectively.Conclusion: We have successfully obtained several clones of functional monoclonal antibodies which can suppress the adherence of esophageal carcinoma cells with lung micro-vascular endothelia, and 2 of the clones show promise in anti metastasis therapy of esophageal carcinoma.

国家自然科学基金重点项目（30230150）；国家重点基础研究发展规划 （973计划)项目 (2002CB513106）