CAJ | 학술논문

目的探讨急性髓系白血病( AML)细胞中硫氧还蛋白(Trx)的表达及其与AML患者临床特点及预后的相关性,观察Trx抑制剂二酰胺体外诱导白血病细胞凋亡的作用,为临床治疗AML提供一定的理论依据.方法选择20例AML患者和10名健康人,抽取其骨髓,分离单个核细胞.用Westernblot法分析单个核细胞中Trx的表达,应用四甲基偶氮唑蓝比色(MTT)法检测不同浓度二酰胺对单个核细胞增殖的影响,应用形态学、琼脂糖凝胶电泳观察二酰胺对细胞凋亡的诱导作用.结果 AML患者Trx的表达率为75 ％(15/20),在健康人中不表达.Trx与患者的白细胞计数显著相关(x2=9.375,P＜ 0.05).MTT法检测显示二酰胺对细胞具有细胞毒作用,且呈时间和剂量依赖性,24、48和72 h的IC50值分别为98.26、47.53、8.34 mg/ml.形态学观察二酰胺作用于AML细胞后可见凋亡小体,琼脂糖凝胶电泳观察可见典型的DNA梯形条带出现.结论 AML细胞Trx含量明显高于健康人,二酰胺干预后AML细胞生长受抑甚至发生凋亡,其可能成为AML治疗的新药.
목적 탐토급성수계백혈병( AML)세포중류양환단백(Trx)적표체급기여AML환자림상특점급예후적상관성,관찰Trx억제제이선알체외유도백혈병세포조망적작용,위림상치료AML제공일정적이론의거.방법 선택20례AML환자화10명건강인,추취기골수,분리단개핵세포.용Westernblot법분석단개핵세포중Trx적표체,응용사갑기우담서람비색(MTT)법검측불동농도이선알대단개핵세포증식적영향,응용형태학、경지당응효전영관찰이선알대세포조망적유도작용.결과 AML환자Trx적표체솔위75 ％(15/20),재건강인중불표체.Trx여환자적백세포계수현저상관(x2=9.375,P＜ 0.05).MTT법검측현시이선알대세포구유세포독작용,차정시간화제량의뢰성,24、48화72 h적IC50치분별위98.26、47.53、8.34 mg/ml.형태학관찰이선알작용우AML세포후가견조망소체,경지당응효전영관찰가견전형적DNA제형조대출현.결론 AML세포Trx함량명현고우건강인,이선알간예후AML세포생장수억심지발생조망,기가능성위AML치료적신약.
Objective To explore the expression level of thioredoxin (Trx) in acute myeloid leukemia (AML) cells,and its association with clinical features and prognosis,as well as the effects of diamide,an inhibitor of Trx,in inducing AML cells apoptosis.Methods Expression of Trx on AML cells and fresh bone marrow cells from healthy adults were analyzed by Western-blotting. The inhibition of proliferation was measured by MTT assay.The anti-leukemia effect of diamide was observed by morphology and agarose gel electrophoresis.Results 75 ％ (15/20) of AML patients expressed Trx,while no expression was observed in control group.Higher expression level of Trx was associated with higher WBC counts (x2 =9.375,P ＜ 0.05),which suggested that overexpression was associated with leukemogenesis.The inhibition of diamide on AML cells showed time and dose dependent by MTT assay.The IC50 values of diamide at 24 h,48 h and 72 h were 98.26 mg/ml,47.53 mg/ml and 8.34 mg/ml,respectively.After AML cells were treated with diamide,the apoptotic body appeared by morphology,and the typical DNA “ladder” bands were confirmed by agarose gel electrophoresis.Conclusion Trx expression level in AML cells is significantly higher than that of control group.Diamide inhibites the proliferation of AML cells by inducing apoptosis,which might be a potential agent for AML.