In vitro killing effect of recombinant perforin peptides on liver cancer cell / 医学研究生学报

ABSTRACT

Objectives:To investigate in vitro tumor killing effect of recombinant human perforin C terminal truncated 125 amino acid polypeptide (rhPFP C) and N terminal truncated 118 amino acid polypeptide (rhPFP N, 22 139aa) using liver cancer cell line SMMC 7721 as target cells. Methods:Recombinant human PFP C and PFP N peptide were expressed by E.coli in fusion form with glutathione S transferase(GST), and were purified by affinity chromatography with glutathione agarose. Liver cancer cell line SMMC 7721 was treated with fusion proteins in different concentrations for 24 h before surviving cells were measured with microscopy and colorimetric MTT assay. Results:After treatment with rhPFP C or rhPFP N, SMMC 7721 cell membrane damaged, appeared lysis. In MTT assay, optical density values in test group were significantly lower than those in control group. At concentration of 2.5 ?g/ml, the killing activity of rhPFP C and rhPFP N were 33.38% and 5.90% respectively. Conclusions:Both rhPFP C and rhPFP N showed obvious killing effect on liver cancer cell line SMMC 7721.The activity of rhPFP C was great higher than that of rhPFP N.