Abstract The purpose of this study was to elucidate the apoptosis,apoptotic pathway of HL-60 cells induced by proteasome inhibitor MG132 and its effect on allogeneic mixed lymphocyte reaction.Apoptosis of HL-60 cells was detected by flow cytometry,the expression of P21,P27 and P53 proteins in HL-60 cells treated with MG132 was assayed by Western blot.The HL-60 cells were treated with 1 μnol/L MG132 for 48 h,and irradiated by 75 Gy of 60 Co γ-ray,but their antigenecity was preserved.The effect of irradiated HL-60 cells treated with MG132 on proliferation of peripheral blood mononuclear cells (PBMNC) was measured by CCK-8 method.The results showed that the apoptotic rate of MG132-treated HL-60 cells increased in dose-and time-dependent manner.No significant changes in MG132-induced apoptosis were observed after inhibiting caspase-8 and caspase-9 pathway.The expression of P21 and P27 protein increased after treatment of HL-60 cells with MG132.CCK-8 test showed that HL-60 cells induced with low-dose of MG132 displayed the enhancing effect on proliferation of PBMNC.It is concluded that high dose of MG132 can induce the apoptosis of HL-60 cells,and has direct killing effect on HL-60 cells,but this inducing apoptotic effect on HL-60 cells can not be realized through caspase-8 and caspase-9 pathway.The P21 and P27 protein may be involved in MG132 induced HL-60 cell apoptosis.Low dose of MG132 promotes the proliferation of PBMNC in healthy individials and enhance the immurnity of organism.%本研究探讨蛋白酶体抑制剂MG132诱导HL-60细胞的凋亡、凋亡途径及其对混合淋巴细胞反应的作用.流式细胞术检测MG132诱导HL-60细胞凋亡、阻断caspase-8和caspase-9通路、MG132诱导HL-60细胞凋亡的变化;Western blot检测MG132作用于HL-60细胞后P21、P27和P53蛋白的表达;应用75 Gy照射经MG132作用的HL-60细胞,然后用CCK-8检测HL-60细胞对健康人单个核细胞的增殖作用.结果表明:大剂量MG132可诱导HL-60细胞凋亡;阻断caspase-8和caspase-9通路后MG132诱导HL-60细胞的凋亡无明显变化,P21蛋白、P27蛋白在MG132作用于HL-60细胞后表达升高,小剂量MG132诱导的HL-60细胞对健康人单个核细胞有增殖作用.结论:大剂量MG132诱导HL-60细胞凋亡,对HL-60细胞有直接杀灭作用,MG132诱导的HL-60细胞凋亡不通过caqspase-8和caspase-9途径,P21和P27蛋白可能参与了MG132诱导的HL-60细胞凋亡,小剂量MG132促进健康人单个核细胞的增殖作用,提高机体的免疫性.
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