重组灵芝免疫调节蛋白抑制HL60细胞增殖及诱导凋亡的研究

Inhibiting and apoptosis-inducing effects of recombinant Ganoderma Lucidum immunoregulatory protein on HL60 cells

目的:探讨重组灵芝免疫调节蛋白(rLZ-8)对HL60细胞抑制作用及诱导凋亡的研究。方法:采用MTT法检测rLZ-8对HL60细胞的体外杀伤作用;通过Annexin V/PI双染检测细胞凋亡率;应用钙离子荧光探针Fluo-3/AM染色,检测rLZ-8作用HL60细胞后细胞内钙离子浓度变化;分光光度法测定细胞caspase-3活性。结果:rLZ-8可明显抑制HL60细胞增殖,随着rLZ-8浓度增加,细胞凋亡率也增加;激光共聚焦检测细胞内钙离子水平明显升高,并且细胞内caspase-3活性增高。结论:rLZ-8可诱导HL60细胞发生凋亡,细胞内钙超载及caspase-3活性增高可能是rLZ-8诱导HL60细胞凋亡的途径之一。

Objective：To investigate the inhibiting and apoptosis-effects inducing of the recombinant Ganoderma Lucidum immunoregulatory protein（rLZ-8） on HL60 cells.Methods：HL60 cells were cultured and the inhibited rate of HL60 cells was measured by means of MTT assay.Apoptosis rate was detected with annexinV/PI.The Calcium ion level was analyzed by Fluo-3/AM stain.Caspase-3 activity was assessed by caspase-3 colorimetric assay.Results：The experiments indicated that rLZ-8 could induce the HL60 cells apoptosis.The calcium ion level was increased and caspase-3 activity elevated in HL60 cells after treated by rLZ-8.Conclusion：rLZ-8 could inhibit the proliferation and induce apoptosis of HL60 cells efficiently.The mechanism may be related to increased calcium ion level and elevation of caspase-3 activity.