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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 study the changes of miRNA expression profile after leukemia cell apoptosis induced by norcantharidin (CA) and to search differentially expressed miRNAs. Methods K562 cells were treated with different concentrations of norcated CA. Cell growth inhibition rates were measured by MTT assay and the effective drug concentration and time were screened. The intracellular miRNAs and microarray hybridization were used to screen the drug effects Post-intracellular changes in miRNA; real-time quantitative PCR to verify microarray hybridization results. Results The effect of 20 mg.L-1 of CA for 36 h was the ideal drug concentration and duration of action. More than 160 miRNAs with more than 120 miRNAs were screened by miRNA microarray hybridization. Further analysis by real-time quantitative PCR It was found that the expression of miR-193B, miR-511, miR-212 and the like were decreased, while the expression of miR-769, miR-125A and miR-100 was significantly increased. Conclusion Norfloxacin can induce the apoptosis of leukemia cells and induce the changes of many miRNAs in the cells.