Effect of granulysin (GLS) and IL-12 genes' expression products on proliferation and apoptosis of melanoma B16 cell / 重庆医科大学学报

ABSTRACT

Objective:To dectect the effect of granulysin and IL-12 genes'expression products on proliferation and apoptosis of melanoma B16 cell in vitro.Methods:Co-expression plasmid including granulysin peptide and murine interleukin 12(mIL-12)genes was transfected into melanoma B16 cell with Lipofectamin TM2000 and its expression products were detected by RT-PCR.Growth suppression was detected with MTT colorimetric assay,and cell apoptotic alterations were evaluated by Hoechst 33258 staining,AO/EB staining,and Annexin V-FITC flow cytometry(FCM).Results:GLS peptite and IL-12 genes could be expressed in B16 cells.Expression products inhibited the proliferation of melanoma cells under MTT observaton.Cells apoptosis with nuclear chromatin condensation,fragmentation and cell membrane change were observed under Hoechst 33258 staining and AO/EB staining.FCM analysis showed the apoptotic rates in test group was 21.02%,which was higher than that in control in control group(15.57%).Conclusion:Expression products of granulysin and mIL-12 genes can not only inhibit proliferation but also induce apoptosis of murine melanoma cell line B16 in vitro.