PTD4-GFP-Apoptin 融合蛋白对白血病细胞增殖与凋亡的影响

目的：研究 PTD4-GFP-Apoptin 融合蛋白对不同类型白血病细胞的增殖抑制及诱导凋亡效应。方法用基因工程技术构建含有 PTD4-GFP-Apoptin 目的基因的重组载体，表达 PTD4-GFP-Apoptin融合蛋白，用四甲基偶氮唑盐比色法（MTT）检测其对白血病细胞增殖的影响，用流式细胞术（FCM）检测其对白血病细胞凋亡的影响。结果 PTD4-GFP-Apoptin 对不同类型的白血病细胞都有不同程度的增殖抑制效应，且抑制效应与作用的时间和浓度呈正相关；PTD4-GFP-Apoptin 对急性髓系白血病细胞株HL-60有诱导凋亡作用，其凋亡率与 PBS 对照组细胞相比差异有统计学意义（P＜0.05）。结论 PTD4能够携带大分子蛋白质穿过细胞膜，PTD4-GFP-Apoptin 在体外对多种白血病细胞有抑制增殖的作用；Apoptin 可诱导白血病细胞凋亡，而正常细胞不受影响，可能成为临床治疗白血病的新型药物。

Effect of PTD4-GFP-Apoptin on proliferation and apoptosis in leukemia cells

Objective To investigate the effects of PTD4-GFP-Apoptin protein on proliferation inhibition and apoptosis-inducing of different types of leukemia cells. Methods Genetic engineering was used to restructure a carrier containing PTD4-GFP-Apoptin gene, and MTT was applied to detect the expressed PTD4-GFP-Apoptin fusion protein and its effect on the leukemia cell proliferation. Flow cytometry (FCM) was used to detect the effects on cell apoptosis. Results MTT cell proliferation inhibitory experiment showed that PTD4-GFP-Apoptin had different degree of proliferation inhibition on different types of leukemia cells;furthermore, the inhibitory effect presented positive correlation with time and concentration. FCM showed that PTD4-GFP-Apoptin had apoptosis-inducing effect on HL-60 cells, and the apoptotic rate had significant difference compared with the control group (P <0.05). Conclusions PTD4 can carry large proteins to penetrate the cell membrane, and PTD4-GFP-Apoptin may produce the inhibiting proliferation in vitro for a variety of leukemia cells. Apoptin can induce tumor cell apoptosis without affecting normal cells, which might become a new agent for the clinical treatment of leukemia.