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,P〈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,P〈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。
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 transfeeted into PC-3 cells by the lipofectin. Transfeeted 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 monoelonal antibody on complement mediated cytotoxieity 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,P〈0.05). At the same concentration of sp22 and anti-CD59 monoclonal antibody, the lysis rate of transfected PC3 cells in sp22 group was obviously higher than that in anti CD59 monoclonal antibody group (q= 7. 805-12. 280 ,P〈0.05). Conclusion sp22 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.