去甲斑蝥素致凋亡K562细胞中miRNA表达的变化

Changes of miRNA Expression of Norcantharidin-Induced Apoptotic K562 Cells

目的研究去甲斑蝥素(CA)致白血病细胞凋亡后miRNA表达谱的变化,寻找差异表达的miRNA。方法应用不同质量浓度的去甲CA处理K562细胞,四甲基偶氮唑盐法检测其细胞生长抑制率,筛选有效的药物质量浓度和作用时间;提取细胞内miRNA,微阵列杂交技术筛选药物作用后细胞内发生变化的miRNA;实时定量PCR技术验证微阵列杂交结果。结果 20 mg.L-1的去甲CA作用36 h为理想的药物浓度和作用时间;miRNA微阵列杂交筛选出表达升高的miRNA 160多种,降低的miRNA 120多种;实时定量PCR进一步分析发现miR-193B、miR-511、miR-212等的表达降低,而miR-769、miR-125A、miR-100等的表达显著升高。结论去甲CA可诱发白血病细胞凋亡,并引起细胞内多种miRNA的改变。

Objective To identify the differentially expressed miRNA,and to investigate the variation of the miRNA expression profile after leukemia cell apoptosis induced by norcantharidin. Methods Growth inhibition ratio of K562 cell was detected by 3-（4,5-Dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide after being treated by different concentration of norcantharidin.Effective drug concentration and treating time were screened.miRNA was extracted from the cell and microarray analysis was used to screen miRNA which showed different expression profiles after being treated with norcantharidin.Real-time polymerase chain reaction was used to validate the selected miRNA. Results The effective drug concentration was 20 mg·L-1 while the treating time was 36 h.There were more than 160 miRNA up-regulated and more than 120 miRNA down-regulated detected by micoarray analysis,and real-time polymerase chain reaction results showed that the expressions of miR-193B,miR-511,miR-212,and so on were decreased,while the expressions of miR-769,miR-125A,miR-100,and so on were increased. Conclusions Norcantharidin can induce K562 cell apoptosis and change the miRNA expression profile.