Objective: To investigate the effect of thrombospondin-1 (TSP-1) on apoptosis of human megakaryocytic leukemia cell line Meg-01 and its possible mechanism.
Methods: The expression of CD36 antigen in Meg-01 cells was detected by flow cytometry and immunocytochemistry. Meg-01 cells were cultured for 48 hours with TSP-1 and CD36 antibody FA6-152 at different concentrations. The early apoptosis and activity of caspase-3 were detected by flow cytometry. The effect of TSP-1 on the growth and differentiation of megakaryocytes was investigated by cell counting and CFU-MK culture.
Results: The flow cytometry and immunocytochemistry showed that CD36 antigen was expressed on the surface of Meg-01 cells. TSP-1 (5 μg/ml) inhibited the growth of Meg-01 cells, but had unobvious effect on M-07e cells. After addition of CD36 antibody FA6-152 (5, 10, and 25 μg/ml), the inhibition effect of TSP-1 was significantly reduced. TSP-1 (2.5, 5, and 7.5 μg/ml) increased the positive expression of Annexin V (P<0.01) and caspase-3 activity (P<0.01), which indicated that TSP-1 had a significant effect on inducing apoptosis. After addition of CD36 antibody FA6-152 (25 μg/ml), the apoptosis induced by TSP-1 in Meg-01 cells was significantly reduced. TSP-1 (5, 10, and 25 μg/ml) could significantly inhibit the formation of CFU-MK in mouse bone marrow cells, while β-TG could not. CD36 antibody FA6-152 (25 μg/ml) could significantly reduce the inhibition of TSP-1 on CFU-MK.
Conclusion: TSP-1 may induce apoptosis of megakaryocytic leukemia cell line Meg-01 cells via CD36/caspase-3, which provides a potential new drug development and treatment target for clinical treatment of megakaryocytic leukemia.
题目: TSP-1通过CD36/Caspase-3诱导巨核细胞白血病细胞系 Meg-01凋亡的研究.
目的: 探讨血小板反应蛋白-1（TSP-1）对人巨核细胞白血病细胞系Meg-01凋亡的影响及可能的机制。.
方法: 应用流式细胞术和免疫细胞化学染色检测Meg-01细胞CD36抗原的表达；采用不同浓度的TSP-1和CD36抗体FA6-152作用于Meg-01细胞，培养48 h，应用流式细胞术检测细胞的早期凋亡和凋亡蛋白酶Caspase-3的活性；采用细胞计数和小鼠巨核细胞集落（CFU-MK）培养探究TSP-1对巨核细胞生长分化的影响。.
结果: 流式细胞术和免疫细胞化学染色结果均显示，Meg-01细胞表面有CD36抗原的表达。TSP-1（5 μg/ml）对Meg-01细胞的生长起到抑制作用，而对CD36-的M-07e细胞作用不明显；加入CD36抗体FA6-152（5、10和25 μg/ml）之后，TSP-1的抑制作用明显减弱。TSP-1（2.5、5和7.5 μg/ml）增加Annexin V的阳性表达（P<0.01），并激活Caspase-3（P<0.01），表明TSP-1诱导细胞凋亡的作用显著；加入CD36抗体FA6-152（25 μg/ml）之后，TSP-1诱导Meg-01细胞凋亡的作用明显减弱。TSP-1（5、10和25 μg/ml）对小鼠骨髓细胞的CFU-MK形成也有明显的抑制作用，而β-TG则没有这种作用；CD36抗体FA6-152 （25 μg/ml）可明显减弱TSP-1对CFU-MK的抑制作用。.
结论: TSP-1有可能通过CD36/Caspase-3诱导巨核细胞白血病细胞系Meg-01细胞凋亡，这为临床上治疗巨核细胞白血病提供了潜在的新药开发研究靶点。.
Keywords: CD36; Meg-01; apoptosis; megakaryocytic leukemia; thrombospondin-1.