三氧化二砷对人骨髓增生异常综合征细胞株MUTZ-1细胞的作用

Inhibition effect of arsenic trioxide on the growth of human MDS cell line MUTZ-1 cells

目的 :研究三氧化二砷 (AS2 O3 )在体外对人骨髓增生异常综合征难治性贫血伴原始细胞过多型 (MDS-RAEB)细胞株 MUTZ- 1细胞增殖的影响 ,并探讨其可能的作用机制。方法 :采用细胞形态学、MTT法、DNA凝胶电泳、RT- PCR、诱导分化检测、流式细胞术等多参数分析。结果 :1低浓度 AS2 O3 (0 .0 5～ 0 .2 5μmol/ L )对 MUTZ-1细胞无明显生长抑制作用 ,细胞培养 2周后 ,细胞阳性表面抗原 CD38、CD7、CD10、HLA- DR表达明显下降 (P<0 .0 5 ) ,粒系分化抗原 CD11b无明显表达增多 (P>0 .0 5 )。 2 AS2 O3 1.0～ 2 0 .0 μmol/ L对 MUTZ- 1细胞有凋亡作用 ,且呈浓度依赖 (r=- 0 .999,P<0 .0 5 )。bcl- 2 m RNA随 AS2 O3 浓度增加表达明显下调 (P<0 .0 5 ) ,bcl- 2 / bax比例明显下降 (P<0 .0 5 )。结论 :0 .0 5～ 0 .2 5 μmol/ L AS2 O3 对 MUTZ- 1细胞可能具有部分分化作用 ;1.0～ 2 0 .0μmol/ L AS2 O3 主要通过诱导凋亡使 MUTZ- 1细胞死亡。

Objective: To investigate the inhibition effect of arsenic trioxide (AS_2O_3) on the growth of human MDS-RAEB cell line MUTZ-1 cells and to explore the possible cellular and molecular mechanisms. Methods: The apoptosis and differentiation of MUTZ-1 cells induced by AS_2O_3 solution of different concentrations were studied with cell morphology,MTT,DNA fragmentation assay,RT-PCR,Nitroblue tetrazolium (NBT) reduction method and flow cytometry. Results: ①Low concentration of AS_2O_3 (0.05～0.25 μmol/L) had no marked growth inhibition effect on MUTZ-1 cells; after 14 d treatment,it down-regulated the expression of positive cell differentiation antigens CD38,CD7,CD10,HLA-DR (P<0.05),but did not up-regulate the expression of CD11b(P>0.05). ②After treatment with 1.0～20.0 μmol/L of AS_2O_3,MUTZ-1 cells presented typical features of apoptosis with a dose dependent manner(r=-0.999,P<0.05). The expression of bcl-2 mRNA and the ration of bcl-2/bax were decreased after AS_2O_3 treatment(P<0.05). Conclusion: Low concentration of AS_2O_3 may have partial differentiation inducement on MUTZ-1 cells. With a certain range of dose (1.0～20.0 μmol/L), AS_2O_3 can induce apoptosis of MUTZ-1 cells. AS_2O_3 can significantly down-regulate bcl-2 and it might be one of the mechanisms of AS_2O_3 treatment.