目的观察噬菌体肽库筛选的短肽(SP22)对前列腺癌细胞高表达的正常或突变CD59分子活性位点的封闭效果。方法将正常和突变CD59基因、pIRES空质粒分别转染PC-3细胞;流式细胞术检测细胞表面CD59分子的表达;RT-PCR检测CD59基因的mRNA表达;补体溶解试验观察SP22对补体介导的PC-3细胞溶解的影响。结果CD59基因成功转染并表达;wPC-3细胞(转染正常CD59基因)和mPC-3细胞(转染突变CD59基因)内CD59 mRNA表达水平显著高于pPC-3细胞(转染空质粒)(P<0.01);SP22使3种细胞的溶解率明显提高(P<0.01),但升高的模式和幅度显著不同。结论SP22不能封闭突变CD59分子的补体结合位点,但可与PC-3细胞表面的正常CD59分子高效结合并中和其补体抑制活性,进而显著增强补体对PC-3细胞的溶解。
Objective To investigate the effects of a docosapeptide(SP22) screened by peptide library on the blockade of activity site of normal or mutant CD59 molecules highly expressed on prostate cancer cells.Methods the pIRES vectors carrying wild-type CD59 gene(wCD59) or mutant CD59 gene(mCD59) and the empty vectors pIRES were transfected into PC-3 cells respectively.The expression of CD59 on PC-3 cells was detected by fluorescence activated cell sorter(FACS),while the level of CD59 mRNA was assayed by semiquantit...