ABSTRACT
Objective To investigate and detect the immunity characteristics of a conserved sequence including 30 amino residues which is located in the C terminal of HPVL1 .Methods The immune model of mouse was establish with a polypeptide synthetized based on this sequence .The amount of CD3+ ,CD3+CD4+ or CD3+CD8+ lymphocyte of the mouse spleen was detected by Flow cy-tometry ,then the value of CD4+ /CD8+ were calculated .The cell proliferation was detected by MTT assay .Sample was took from the cell supernatant to detect the content of IL-4 and IFN-γby double antibody sandwich ELISA .Results (1)The amount of CD3+CD8+ lymphocytes and the value of CD4+ /CD8+ were significantly increased than control group(P0 .05) .(3)The level of IL 4 were significantly higher than that of control group (P0 .05) .Conclusion The results showed that it was affirmative that the polypeptides induced humoral immunity ,which was worth to be furtherly studied as a preventive vaccine .But its cell immunogenicity is weak ,and the attenmpt to induce the response related to the cell immunogenicity was failed ,and the immunogenicity of the peptide remains to be improved .
ABSTRACT
Aim To study the effects of polyamine analogue CPENSpm on the human lung cancer line A549 in cell proliferation and apoptosis.Methods MTS was used to assay the cell proliferation,chemical analysis methods were used to determine the activities of enzymes in the polyamine metabolism,HPLC was performed to assay the intracellular concentration of polyamines,Sub-G1 and DNA fragmentation assays were used to determine the cell apoptosis.Results Treating A549 lung cancer cells by CPENSpm resulted in:①cell-growth inhibition and cell apoptosis;②inhibition of ODC(key enzyme in polyamine synthetic pathway)and activation of SSAT and SMO(key enzymes in polyamine catabolism);③great decrease of intracellular polyamine concentrations.MDL72527,the SMO inhibitor,can antagonize the effect of CPENSpm on inhibiting the proliferation of A549 cells.Conclusion CPENSpm inhibits proliferation and induces apoptosis of human A549 lung cancer cell line by interfering the polyamine metabolism,depleting intracellular polyamine contents that are need by quick-growth of cancer cells and inducing production of H2O2.