六亚甲基二乙酰胺对K562细胞作用的实验研究

Research of hexamethylene bisacetamide’s effect on K562 cells in vitro

目的:研究六亚甲基二乙酰胺(HM-BA)体外对K562细胞的抑制增殖、诱导凋亡和促进分化的作用。方法:MTT法检测不同浓度HMBA对于K562细胞的增殖抑制率;流式细胞仪(FCM)检测HMBA对于K562细胞周期分布的影响;AnnexinV/PI染色方法检测HM-BA诱导K562细胞凋亡的作用;通过瑞氏染色、联苯胺染色和FCM检测分化抗原研究K562细胞分化情况。结果:HMBA可以抑制K562细胞增殖,1、2、3和4mmol/L对K562细胞增殖抑制率分别为21.97%、36.05%、41.56%和47.59%;HMBA可以阻滞K562细胞周期,主要表现为G0/G1期的阻滞;HMBA诱导K562细胞凋亡的作用不明显,4mmol/L的HMBA对K562细胞诱导凋亡率<5%;HMBA处理后,瑞氏染色显示K562细胞有一定程度的成熟表现,联苯胺染色基本为阴性;K562细胞膜表面CD11b、CD14、CD68和胞质内溶菌酶抗原的表达在HMBA处理前后均为阴性。结论:HM-BA可以抑制K562细胞的增殖,提示具有一定的治疗作用;HMBA对K562的作用机制和阻滞细胞周期有关,诱导凋亡不是主要作用机制;HMBA有促进K562细胞分化的迹象,但分化方向和机制有待进一步研究。

OBJECTIVE： To investigate the effect of hexamethylene bisacetamide （HMBA） on K562 cells in vitro. METHODS： MTT colorimetric assay was used to measure HMBA＇s inhibitory ability to K562 cells＇ proliferation. Flow Cytometry （FCM） was used to analyse the changed cell cycle＇s distribution of K562 cells induced by HMBA. Double staining of Annexin V/PI was used to detect the apoptosis of K562 cells. The differentiation of K562 cells induced by HMBA was evaluated by Giemsa staining and benzidine staining. At the same time, the antigens of differentiation were detected by FCM. RESULTS： In vitro, HMBA could inhibit the proliferation of K562 cells, 1, 2, 3 and 4 mmol/L of HMBA＇s inhibition ratio to K562 cells correspondingly was 21.97%, 36.05%, 41.56% and 47.59% ; commit K562 cells to G0/G1 phase arrest, and induce K562 cells trivial apoptosis 4 mmol/L of HMBA＇s inducing apoptisis ratio to K562 cells was less than 5%. After treatment of HMBA, K562 cells seemed to show differentiation, from the result of Giemsa staining. However, the result of benzidine was negative. At the same time, K562 cells didn＇t show observable expression of antigen markers of differentiated granulocyte or monocyte-rnacrophage. CONCLUSIONS： HMBA can inhibit the proliferation of K562 cells, which indicates HMBA has potential of treating chronic myelogenous leukemia. To understand the precise mechanism, more research is needed.