2-甲氧基雌二醇对K562细胞L-选择素及其基因表达的影响  

Effects of 2-methoxyl estradiol on the expression of L-selectin protein and its mRNA in K562 cells

作　　者：李栋梁[1] 潘崚[1] 

机构地区：[1]河北医科大学第二医院血液病科,石家庄050000

出　　处：《解放军医学杂志》2007年第12期1270-1273,共4页Medical Journal of Chinese People's Liberation Army

基　　金：河北省科学技术研究与发展指导计划资助项目(072761183)

摘　　要：目的观察2-甲氧基雌二醇(2-ME)对慢性粒细胞白血病细胞系K562细胞L-选择素表达的影响,并通过分析L-选择素与bcr/abl融合基因表达的关系探讨2-ME的作用机制。方法实验分3组。①对照组:培养基中不含2-ME;②实验组:分别用1、2、4、8、16μmol/L的2-ME处理K562细胞36h,观察各项指标的变化;③阴性对照组:培养基中以无RNase水代替K562细胞。用流式细胞术(FCM)检测K562细胞L-选择素蛋白表达;半定量RT-PCR检测L-选择素mRNA的表达;SYBRGreenⅠ实时荧光定量RT-PCR检测bcr/abl融合基因的mRNA表达。结果不同浓度的2-ME与K562细胞作用36h后,L-选择素蛋白及其mRNA的表达量均呈剂量依赖性增加,与对照组比较均有显著性差异(P<0.05,P<0.01);L-选择素mRNA与其蛋白表达之间呈正相关(r=0.758,P=0.004)。L-选择素mRNA与bcr/abl融合基因mRNA之间呈负相关(r=-0.968,P=0.001)。结论2-ME可能通过抑制K562细胞bcr/abl融合基因的表达,增加L-选择素mRNA及其蛋白表达水平,改善或恢复K562细胞的黏附缺陷。Objective To investigate the effects of 2-methoxyl estradiol（2-ME）on the expression of L-selectin and its mRNA and the relationship between L-selectin and bcr/abl fusion gene expression in chronic myelocytic leukemia（CML）K562 cells.Methods The cultured K562 cells were divided into three groups.The control group,K562 cells were cultured without 2-ME treatment.The experimental group,K562 cells were cultured in the medium containing different concentrations of 2-ME（1,2,4,8 and 16μmol/L）for 36h.The negative control group,K562 cells were replaced by water without RNase in the medium containing different concentrations of 2-ME for 36h.The expression of L-selectin protein in K562 cells was determined by flow cytometry（FCM）method.The expression of L-selectin mRNA in K562 cells was detected by half-quantitative reverse transcription-polymerase chain reaction（RT-PCR）.The bcr/abl fusion gene expression in K562 cells was detected by quantitative SYBR Green Ⅰ real-time RT-PCR.Results After being treated with 2-ME at different concentrations for 36 hours,the expression of L-selectin protein and its mRNA increased in K562 cells in a concentration-dependent manner,and the expression level in the experimental group was higher than that in the control group（P〈0.05,P〈0.01,respectively）.The expression levels of L-selectin mRNA in K562 cells were positively correlated with that of L-selectin protein（r=0.758,P=0.004）.③The expression levels of L-selectin gene were negatively correlated with that of bcr/abl fusion gene expression（r=-0.968,P=0.001）.Conclusion 2-ME may increase the expression of mRNA and protein of L-selectin by down-regulating bcr/abl fusion gene expression,which may improve or restore the aberrant adhesion of K562 cells.

关 键 词：白血病 髓样 慢性 L选择素 细胞黏附分子 

分 类 号：R557.3[医药卫生—血液循环系统疾病]