去甲斑蝥素致凋亡白血病细胞基因表达的变化  

Changes of Gene Expression in Apoptotic Leukemia Cells Induced by Demethylcantharidin

作　　者：李有杰[1] 张帅[1] 王萍玉[1] 高宗华[2] 王秀文[3] 谢书阳[1] 

机构地区：[1]滨州医学院医学分子遗传研究所,山东烟台264003 [2]滨州医学院化学教研室,山东烟台264003 [3]滨州医学院实验医学研究中心,山东烟台264003

出　　处：《实用儿科临床杂志》2010年第21期1662-1664,共3页Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金(30801324);山东省自然科学基金(ZR2009CQ033);山东省教育厅课题(J08LG15)

摘　　要：目的研究去甲斑蝥素(DMC)对慢性粒细胞白血病细胞株K562细胞凋亡的影响,探讨DMC致细胞凋亡后基因表达的变化。方法 20 mg.L-1DMC作用细胞36 h后,倒置显微镜观察细胞形态变化;提取细胞DNA,琼脂糖凝胶电泳检测凋亡细胞DNA改变;提取细胞总RNA,基因芯片技术检测基因表达谱变化;通过反转录(RT)-PCR、ELISA验证芯片部分基因表达变化。采用SPSS13.0软件进行统计学分析。结果 DMC处理K562细胞后,倒置显微镜下可见细胞状态变差,活细胞数目明显减少;DNA电泳后可见明显的阶梯状变化;在利用基因芯片技术检测的21 522个基因转录本中,共筛选出差异表达基因约2 300个,其中超过500个基因的表达明显升高,有350个基因表达显著降低;RT-PCR和ELISA结果显示,Bcl-2、E2F1和E2F3基因表达显著降低,RB1和P53表达显著增高,均与芯片结果一致。结论 DMC可致K562细胞凋亡,并可引起凋亡细胞相关基因表达变化。Objective To explore the effect of demethylcantharidin（DMC） on K562 and the changes of gene expression after cells were treated with DMC.Methods Thirty-six hours after K562 cells were treated with 20 mg·L^-1 DMC,the shape and number of cells were observed by inverted microscopy;the changes of DNA were analyzed by electrophoresis after DNA isolation;mRNA microarrays assessment was used to detect the variation of gene expression after total RNA was isolated from cells;then the expression of several gene was verified by reverse transcription-polymerase chain reaction（RT-PCR） and enzyme-linked immunosorbent assay（ELISA）.The data were analyzed by SPSS 13.0 software.Results After DMC treatment,the shape was changed and the number of alive cells was reduced under inverted microscopy;Clear characteristic DNA ladder zone was showed on agarose gel electrophoresis;mRNA gene microarrays results showed that about 2 300 differentially expressed genes were selected from 21 522 different genes transcripts.The expressions of more than 500 genes were twice higher than the control group,while the expressions of 350 genes were significantly decreased.RT-PCR and ELISA results showed that the expressions of RB1 and P53 were increased significantly in DMC treated group,while the expressions of Bcl-2,E2F1 and E2F3 were significantly decreased.Conclusion DMC can induce leukemia cell apoptosis and variation of relative genes expression in apoptotic cells.

关 键 词：去甲斑蝥素 白血病 细胞凋亡 基因表达 

分 类 号：R285.5[医药卫生—中药学] R733.7[医药卫生—中医学]