烷化溶血磷脂对U266细胞作用的实验研究  

THE EFFECT OF ALKYL-LYSOPHOSPHOLIPID (ET-18-OCH_3) ON U266 CELLS

作　　者：黄春玲 陈志哲[1] 陈君敏[1] 王少元[1] 刘庭波[1] 杨婷[1] 

机构地区：[1]福建医科大学附属协和医院,福建省血液病研究所,福州350001 [2]厦门中山医院临床检验中心,361004

出　　处：《细胞生物学杂志》2003年第6期370-375,共6页Chinese Journal of Cell Biology

摘　　要：为研究烷化溶血磷脂 (ET 1 8 OCH3 )对骨髓瘤细胞系U2 66的体外杀伤作用。通过台盼蓝拒染法和克隆形成率来反映ET 1 8 OCH3 对U2 66细胞的抗增殖效应 ;通过荧光显微镜、电镜观察细胞形态学变化 ,琼脂糖凝胶电泳观察DNA降解片段 ,流式细胞仪作DNA倍体分析来观察ET 1 8 OCH3 诱导U2 66细胞凋亡的作用 ;通过RT PCR检测bcl 2、c mycmRNA水平 ,流式细胞仪检测Bcl 2、C myc蛋白的表达来初步探讨其相关的作用机制。结果显示 ,U2 66细胞经ET 1 8 OCH3 处理后细胞生长明显受抑 ,呈时间和剂量依赖性 ;荧光显微镜 ,电镜下观察均可见凋亡形态学改变 ,7.5μg/mlET 1 8 OCH3 作用 2 4小时后出现典型的DNA降解片段。流式细胞仪检测凋亡率为 1 7.53 % ;RT PCR显示bcl 2、c mycmRNA表达均随ET 1 8 OCH3 作用时间的增加而减弱 ,ET 1 8 OCH3 作用 2 4小时后 ,bcl 2、c mycmRNA分别减少了 86 %、72 % ;流式细胞仪检测Bcl 2蛋白、C myc蛋白分别减少了原来的 1 7%、60 %。研究结果表明 ,ET 1 8 OCH3 对U2 66细胞的生长具有明显的抑制作用 ,并可诱导细胞的凋亡 。To study the anti tumor effects of alkyl lysophospholipid ET 18 OCH 3 (ALP) on U266 cells. The antiproliferative activity of ET 18 OCH 3 was measureed by the inhibitory rate and colony formation assays of U266 cells during the incubation with ET 18 OCH 3. The effect of ET 18 OCH 3 in apoptosis induction was determined by the observation of morphology, agarose gel electrophoresis, and flow cytometry. The expression of bcl 2?c myc mRNA and proteins were detected by RT PCR and flow cytometry in order to understand the mechanism of apoptosis induction. Results showed that ET 18 OCH 3 owned significant cytotoxity on U266 cells and inhibitory effect on colonies formation. These effects were time and dose dependent. ET 18 OCH 3 could also induce time dependent apoptosis, which was confirmed by morphology characteristics and DNA fragments ladder on electrophoretogram. The apoptosis rate in flow cytometry was 17.53%, with no apoptotic cells found in control. RT PCR showed bcl 2 and c myc mRNA decreased by 86% and 72% respectively after U266 cells were incubated with ET 18 OCH 3 for 24h. In the mean time, the percentage of bcl 2?C myc positive cell decreased by 17% and 60%, respectively. These results suggest that ET 18 OCH 3 has significant inhibitory effect on growth of U266 cells, possibly due to its apoptosis induction in U266 cells.

关 键 词：烷化溶血磷脂 U266细胞 多发性骨髓瘤 细胞凋亡 

分 类 号：R73-36[医药卫生—肿瘤]