金钗石斛多糖对髓系白血病细胞WT1基因表达的影响  

Study on effect of dendrobium nobile polysaccharides on expression of WT1 gene in myeloid leukemia cells

作　　者：葛晓军[1] 郑丽梅[2] 王永伦[1] 唐彦萍[3] 

机构地区：[1]遵义医学院第一附属医院检验科,贵州遵义563003 [2]遵义医学院第一附属医院中医科,贵州遵义563003 [3]遵义医学院生物化学与分子生物学教研室,贵州遵义563003

出　　处：《重庆医学》2015年第10期1305-1307,1310,共4页Chongqing medicine

基　　金：国家自然科学基金资助项目(30960509/C190704)

摘　　要：目的探讨金钗石斛多糖对髓系白血病细胞WT1基因表达的影响。方法用CCK8方法检测金钗石斛多糖在3种白血病细胞中的IC50值;3种细胞各分两组,对照组和处理组,对照组细胞保持正常生长,处理组给予金钗石斛多糖刺激。Hoechst33258染色检测两组细胞凋亡情况,用RT-PCR检测WT1表达水平,Western blot检测WT1、P53和BAX蛋白表达。结果金钗石斛多糖在3种白血病细胞中具有相近的IC50值,HL-60、WEHI-3、K562细胞的IC50分别为(110.71±6.49)、(104±48.50)、(96.66±5.10)mg/mL,3种细胞的IC50值比较,差异无统计学意义(P>0.05);处理组凋亡率高于对照组(P<0.05);与对照组比较,处理组WT1基因、蛋白表达水平降低(P<0.05),P53和BAX蛋白表达增高。结论金钗石斛多糖能明显降低WT1蛋白表达水平,且对白血病细胞有一定杀伤作用。Objective To explore the effect of dendrobium nobile polysaccharides on the expression of the WT 1gene in myeloid leukemia cells.Methods The CCK8 assay was used to detect the half maximal inhibitory concentration（IC50）of dendrobium nobile polysaccharides in 3kinds of leukemia cells;the each kind of leukemia cells were divided into the treatment group and the control group.The cells in the control group maintained the normal growth,while which in the treatment group were given the dendrobium nobile polysaccharides stimulation.The Hoechst33258 staining was used to detect the apoptosis situation of the cells in the two groups.The WT l gene expression level was detected by the real-time PCR and the protein expression levels of WT1,53 and BAX were detect the Western blot.Results Dendrobium nobile polysaccharides had the similar IC50 values in 3kinds of myeloid leukemia cells,which were（110.71±6.49）,（104±48.50）,（96.66±5.10）mg/mL respectively,the difference among them had no statistical significance（P〉0.05）;the apoptosis rate of the treatment group was higher than that of the control group（P〈0.05）;the expression levels of WT1 gene and protein in the treatment group were decreased compared with the control group（P〈0.05）,while the expression of P53 and BAX protein was increased.Conclusion Dendrobium nobile polysaccharides can obviously decrease the expression level of WT1 protein,and has a certain killing effect on myeloid leukemia cells.

关 键 词：石斛 多糖 基因 肾母细胞瘤 白血病 

分 类 号：R285[医药卫生—中药学]