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

1)在HEK293细胞和MT1-MMP稳定转染的HEK293之间评价131 I标记的抗MT1-MMP抗体114 F1的细胞毒性和细胞抑制作用。集落形成实验显示两种细胞间无差异，表明抗MT1MMP与131I联合作用不具有细胞抑制作用。而加入MMP抑制剂BB94后，细胞表面MT1-MMP的表达增强，当加入100 μ g抗体时，抗体与细胞的结合亲和力提高了10~20%。2）流式细胞术检测114 F、113 - 5B7和AB815对MT1-MMP的亲和力。结果表明，与114 F或113 - 5B7相比，AB 815具有5至10倍的结合亲和力。结论：3）EB病毒原癌基因LMP1定位于细胞膜上。首先设计了抗LMP 1抗体的建立。但是，LMP1的胞外结构域只有6~7个氨基酸，不足以产生抗体。4）随后，我们将策略转移到寻找LMP1诱导的细胞表面分子上。Mucin1是LMP 1诱导的特异性分子，其作用效果有待于后续实验的进一步研究。

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.