Development of molecular targeting therapy for head and neck cancer by targeting membrane type MMP and EBV antigen

1)评价131I标记的抗MT1-MMP抗体114F1在HEK293细胞和MT1-MMP稳定转染的HEK293细胞间的细胞毒和细胞抑制作用。集落形成实验显示两种细胞间无差异，表明131I抗mt1mmp不具有细胞抑制作用。然而，添加MMP抑制剂BB94可增强细胞表面MT1-MMP的表达，当给予100ug抗体时，抗体与细胞的结合亲和力提高了10% ~ 20%。2)利用流式细胞术重新评估114F、113-5B7和AB815对MT1MMP的亲和力。结果表明，AB815与114F和113-5B7的结合亲和力是前者的5 ~ 10倍。这些结果表明，使用高亲和力抗体和MMP抑制剂可以改善针对MT1-MMP的分子靶向治疗。3) eb病毒原发癌基因LMP1位于细胞膜上。首先设计了LMP1抗体的建立。但LMP1的外结构域有6 ~ 7个氨基酸，不足以产生抗体。4)然后，将策略转换为寻找LMP1诱导的细胞表面分子。Mucin1是LMP1诱导的特异性分子，后续实验需进一步检测抗Mucin1抗体的作用。

1)Cytotoxic and cytostatic effect of Anti-MT1-MMP antibody 114F1 labeled with 131I was evaluated between HEK293 cell and MT1-MMP stable transfected-HEK293. Colony, formation assay showed there was no difference between these two cells, indicating that anti-MT1MMP with 131I did not have cytostatic effect. However, additional BB94, an inhibitor of MMP, enhanced the expression of cell surface MT1-MMP, and binding affinity of antibody to cell was enhanced 10 to 20% when 100ug of antibody was administrated.2)Affinity to MT1MMP was reevaluated among,114F,113-5B7 and AB815 using flow cytometry. It showed that AB815 had 5 to 10 fold binding affinity compared with either 114F or 113-5B7. These, results suggests that molecular targeting therapy against MT1-MMP would be improved by the use of higher affinity antibodies and MMP inhibitors.3)Epstein-Barr virus primary oncogene LMP1 locates on cell membrane. Establishment of antibody to LMP1 was first designed. However, ectodomain of LMP1 was revealed to have 6to 7 amino acids, which is insufficient to raise antibody.4)Then, the strategy was switched to look for the cell surface molecule induced by LMP1. Mucin1 was found to a specific molecule induced by LMP1 and effect, of antibody against-mucin1 should be examined in the successional experiment.