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羟地吗啉对慢性粒细胞白血病K562细胞作用的研究 被引量:3

Effects of Hydroxyl Morpholine Resistance on K562 Cells of Chronic Myelocytic Leukemia
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摘要 目的研究羟地吗啉对慢性粒细胞白血病(CML)K562细胞增殖抑制、诱导分化及凋亡的作用。方法采用血细胞计数法绘制细胞生长曲线;阳性对照药为伊马替尼(imatinib),利用四甲基偶氮唑蓝法(MTT)法测定羟地吗啉对K562细胞增殖的影响;软琼脂克隆法观察细胞增殖能力及羟地吗啉的作用;利用Wright-Giemsa染色法进行形态学分析;利用AO-EB双染法和DNA断裂法检测细胞凋亡机制。应用Western-blot方法从蛋白水平分析羟地吗啉对K562细胞P210bcr/abl蛋白的变化。结果由细胞生长曲线可见,K562细胞具有良好的细胞活力,在传代后第3天进入对数生长期;羟地吗啉、伊马替尼对K562细胞的IC_(50)平均值分别为5.81、596.88 nmol·L-1。软琼脂克隆形成试验显示,21 d可以形成克隆,羟地吗啉的克隆形成抑制率为81.7%,K562细胞对羟地吗啉比较敏感。形态学实验结果显示,羟地吗啉能够诱导K562细胞向正常的细胞分化。AOEB双染法结果显示,羟地吗啉能够诱导K562细胞凋亡。DNA断裂研究表明,羟地吗啉可以诱导K562细胞凋亡产生梯状DNA条带。Western-blot免疫印迹法表明,羟地吗啉可下调P210bcr/abl蛋白的表达。结论羟地吗啉对慢性粒细胞白血病K562细胞的增殖具有抑制作用,羟地吗啉能够诱导K562细胞凋亡及向正常细胞分化。 OBJECTIVE To investigate the inhibitory activity, induced differentiation-inducing activity and apoptosis-inducingac- tivity of hydroxyl morpholine( QDML-01 ) on chronic myeloeytie leukemia cells line K562. METHODS The cell growth curve was drawn based on cell counting method. The IC5o value of QDML-01 and positive control medicine to K562 cells were evaluated by methyl thiazolyl tetrazolium (MTI') assay method. Double soft agar assay method was carried out to study the ability of cell proliferation to determine efficacy of phamacognosy. The pathomorphism was analyed by the Wright-Giemsa staining method. The mechanism of cell apoptosis from morphology and gene level were investigated, by AO-EB double-staining method and DNA breakage test. The effect of QDML-01 on K562 cells from the protein level was determined by Western-blot. RESULTS The growth curves showed the I(562 cells had strong cell vitality. They came into logarithmic phase on the third generation. The MTT assay results showed that the ICs0 values of QDML-01 and imatinib to K562 cells were 5.81 and 596. 88 nmol . L-1. Double soft agar colony formation test showed that clone formed at 21 d and the inhibitory rate of QDML-01 was 81.7%. It indicated that K562 cells were sensitive to QDML-01. Morphology test result showed that QDML-01 induced K562 cells to normal cells. The results of AO-EB double-staining method showed that QDML- 01 induced the apoptosis of K562 cells. The study of DNA breakage test indicated that QDML-01 can induce the apoptosis of K562 cells to produce DNA banding with step-like. Western-blot analysis result suggested that QDML-01 can downregulated the expression of P210bcr/ablprotein. CONCLUSION QDML-01 has the inhibitory activity on chronic myelocytic leukemia cells line K562 by promoting the apoptosis of K562 cells and inducing differentiation to normal cells.
机构地区 辽宁大学药学院
出处 《中国药学杂志》 CAS CSCD 北大核心 2016年第5期368-372,共5页 Chinese Pharmaceutical Journal
基金 沈阳市科技局社会发展计划资助项目(F11-140-9-00)
关键词 羟地吗啉 慢性粒细胞白血病 K562细胞株 抗肿瘤活性 伊马替尼 hydroxyl morpholine chronic myeloeytic leukemia K562 cells line antiproliferation activity imatinib
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