CD59结合短肽对补体介导的前列腺癌细胞溶解影响.docVIP

CD59结合短肽对补体介导的前列腺癌细胞溶解影响 【摘要】 目的 观察CD59结合短肽(sp22)对补体介导高表达人CD59分子的PC3细胞溶解的影响。方法利用脂质体法将携带人野生型CD59基因的pIRES质粒(wCD59pIRES)转染PC3细胞，G418筛选稳定转染的细胞株，荧光免疫组化技术检测细胞表面CD59分子的表达，补体溶解试验观察sp22和抗CD59单克隆抗体对补体介导的高表达CD59分子的PC3细胞溶解的影响。结果 高表达CD59分子的PC3细胞株构建成功;与对照组比较，sp22组转染细胞的溶解率显著升高(F=719.552，q=12.181～79.942，Plt;0.05);相同浓度条件下，sp22组转染细胞的溶解率明显高于抗CD59单克隆抗体组(q=7.805～12.280，Plt;0.05)。结论 sp22可封闭PC3细胞表面高表达CD59分子的补体结合位点，显著增强抗PC3细胞抗体所诱发的补体介导的抗瘤作用。 【关键词】 抗原，CD59;短肽;前列腺肿瘤;补体溶解试验 [ABSTRACT] Objective To investigate the effect of short peptide binding to CD59 molecules on lysis of complementmediated prostate cancer PC3 cells which highly expressed transfected human wildtype CD59 gene. Methods pIRES vectors carrying wildtype CD59 gene (wCD59pIRES) were transfected into PC3 cells by the lipofectin. Transfected PC3 cells were screened by the addition of G418. The expression of CD59 on PC3 cells was detected by fluorescence immunohistochemistry technique. The effect of sp22 or antiCD59 monoclonal antibody on complementmediated cytotoxicity to transfected PC3 cells was evaluated by complement lysis assays. Results PC3 cells highly expressing CD59 molecules were successfully constructed. Compared with the control group, the lysis rate of transfected PC3 cells in sp22 group was increased significantly (F=719.552,q=12.181-79.942,Plt;0.05). At the same concentration of sp22 and antiCD59 monoclonal antibody, the lysis rate of transfected PC3 cells in sp22 group was obviously higher than that in antiCD59 monoclonal antibody group (q=7.805-12.280,Plt;0.05). Conclusionsp22 can effectively block the complementbinding sites of CD59 molecules highly expressed on PC3 cells, therefore, the antitumor effect of the complement stimulated by antiPC3cell antibody can be enhanced. 　　[KEY WORDS] Antigen, CD59; Shortpeptide; Prostatic neoplasms; Complement lysis assays 　　CD59是一种以糖基磷脂酰肌醇(GPI)锚着于细胞膜表面的具有抑制补体效应功能的糖蛋白，通过与C8或C9竞争性结合而抑制膜攻击复合物在细胞膜上的形成，是宿主细胞免受活化的补体攻击的最重要的膜补体调节蛋白(mCRP)。大量研究表明，C