论文部分内容阅读
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 observe the blocking effect of phage peptide library-screened short peptides (SP22) on the active site of normal or mutant CD59 molecules highly expressed in prostate cancer cells. Methods The normal and mutant CD59 genes and empty pIRES plasmids were transfected into PC-3 cells respectively. The expression of CD59 on cell surface was detected by flow cytometry. The expression of CD59 mRNA was detected by RT-PCR. Effect of PC-3 cell lysis. Results The expression of CD59 mRNA in wPC-3 cells (transfected with normal CD59 gene) and mPC-3 cells (transfected with mutant CD59 gene) was significantly higher than that in pPC-3 cells (transfected with empty plasmid) (P <0.01). The dissolution rate of three kinds of cells was significantly increased by SP22 (P <0.01), but the mode and extent of increase were significantly different. Conclusions SP22 can not block the complement binding site of mutant CD59 molecule, but can bind efficiently to normal CD59 molecule on the surface of PC-3 cells and neutralize its complement inhibitory activity, thereby significantly enhancing the complement lysis of PC-3 cells.