Construction Of CD59 Ligand-peptide Eukaryotic Expression System And The Effect On Activity To PC-3 Cells

Objective To construct CD59 Ligand-peptide (sp22) eukaryotic expression system, and investigate the expression and function of CD59 on PC-3 cells, which is aim to futher study the inter-mechanism of sp22 and CD59 as well as using Ligand-peptide to restrain CD59 expressing on PC-3 cells surface.Methods Ligand-peptide of CD59 eukaryotic expression system was constructed.Then recombinants were transfected into PC-3 cells by liposome. The stable transfected clones were screened by the addition of G418; The level of CD59 mRNA were detected by RT-PCR; Western Blot,ELISA methods were used to determine the protein expression; Prepared anti-CD59 polyclonal antibody, cake uterine blue staining method and LDH method were used to observe the influence of sp22 on CD59 molecule; MTT was used to determine the growth-inhibition ratio.Results Ligand-peptide of CD59 eukaryotic expression system had been successfully constructed according to sequence of enzyme digestion analysis and fragment investigation; RT-PCR suggest that sp22 was expressed on spPC-3 cells successfully; Western Blot,ELESA competitive inhibition assays suggest that the expression level of CD59 molecule was reduced on spPC-3 cells than the expression level of CD59 molecule on normal PC-3 cells; Compared with normal PC-3 cells the rate of destruction was enhancement obviously, the differences were statistically significant (P<0.05).The result of MTT suggests that the growth-inhibition ratio increased, the differences were statistically significant (P< 0.05)Conclusions CD59 Ligand-peptide (sp22) can influence CD59 mRNA and protein expression level on spPC-3 cells, and also can decrease CD59 molecule anti-complement activity. Results indicate that expressing CD59 Ligand-peptide (sp22) may be a potential way to treat tumors and to establish the base for tumors immunotherapy.