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  

Apoptosis of Megakaryocytic Leukemia Cell Line Meg-01 Induced by TSP-1 Via CD36/Caspase-3

作　　者：孔惠敏 苏伟青[2] 罗毅[2] 戈慧 李亮[4] 杨默 江千里[1] KONG Hui-Min;SU Wei-Qing;LUO Yi;GE Hui;LI Liang;YANG Mo;JIANG Qian-Li(Department of Hematology,Nanfang Hospital,Southern Medical University,Guangzhou 510515,Guangdong Province,China;Department of Oncology Hematology,Lianjiang People′s Hospital,Zhanjiang 524400,Guangdong Province,China;Department of Pediatrics,Nanfang Hospital,Southern Medical University,Guangzhou 510515,Guangdong Province,China;Research Center,The Seventh Affiliated Hospital,Sun Yat-sen University,Shenzhen 518107,Guangdong Province,China)

机构地区：[1]南方医科大学南方医院血液科,广东广州510515 [2]廉江市人民医院肿瘤血液科,广东湛江524400 [3]南方医科大学南方医院儿科,广东广州510515 [4]中山大学附属第七医院科研中心,广东深圳518107

出　　处：《中国实验血液学杂志》2022年第4期998-1004,共7页Journal of Experimental Hematology

基　　金：国家自然科学基金面上项目(81770116)。

摘　　要：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(5μg/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25μg/ml)之后,TSP-1的抑制作用明显减弱。TSP-1(2.5、5和7.5μg/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(25μg/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25μg/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(25μg/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。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(5μg/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 25μg/ml),the inhibition effect of TSP-1 was significantly reduced.TSP-1(2.5,5,and 7.5μg/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(25μg/ml),the apoptosis induced by TSP-1 in Meg-01 cells was significantly reduced.TSP-1(5,10,and 25μg/ml)could significantly inhibit the formation of CFU-MK in mouse bone marrow cells,whileβ-TG could not.CD36 antibody FA6-152(25μg/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.

关 键 词：血小板反应蛋白-1 巨核细胞白血病 Meg-01 CD36 凋亡 

分 类 号：R733.7[医药卫生—肿瘤]