烷化溶血磷脂ET-18-OCH3对K562细胞作用的研究

THE EFFECT OF ALKYL-LYSOPHOSPHOLIPID ET-18-OCH3 ON K562 CELL

为研究烷化溶血磷脂ET-18-OCH3(ALP)的抗白血病效果,本文以K562细胞为研究对象,通过台盼蓝拒染法测定ALP作用后K562细胞的生长抑制率和生长曲线;甲基纤维素半固体培养法测定克隆原细胞的存活率;流式细胞仪检测K562细胞P210蛋白表达;RT-PCR半定量法测定细胞的bcr-abl mRAN;采用流式细胞仪进行DNA分析以及电镜观察细胞形态学改变。结果显示,K562细胞经ALP处理后细胞生长明显受抑制,呈作用时间和剂量的依赖性,IC_(50)为31.6(24),22.3(48h),14.8(72h)μg/ml;细胞增殖速度显著降低,克隆原细胞存活曲线呈指数型,而正常对照组细胞的CFU-GM则未受影响;ALP还可使K562细胞P210及bcr-abl mRNA水平下调,并有诱导细胞凋亡的作用。说明ALP对K562细胞生长具有明显抑制作用,并有诱导细胞凋亡作用,提示ALP具有一定的抗白血病效应。

To study the anti-leukemic effects of alkyl-lysophospholipid ET-18-OCH3(ALP)on K562 cell, cell inhibitory rate and growthcurve were determined by trypan blue dye exclusion. CFU-K562 cells were cultured in 0. 8% methy[cellulose. P210 was measured by flow cytometry. Cellular hcr-abl mRNA was detected by RT-PCR seni-quantitative analysis. Cells apoptosis was measured by flow cytometry and observed by electron microscopy. The result showed that the growth of K562 cell was significantly inhibited as treated with ALP, showing time-quantity dependent effects. The IC50 to K562 cell of ALP was 31.6, 22.3,14. 8μg/ml after in vitro treatment for 24,48, 72h, respectively. The cell proliferation markedly decreased in K562 cell. Coloning forming efficiency assay suggested a /ig effect dose-survival curve of exponential type. ALP also decreased cellular bcr-abl mRNA and P210 expression and induced K562 cell apoptosis. These results suggest that ALP has significant inhibitory effect on growth of K562 cell and also induces apoptosis in K562 cell.