斑蝥素对乳腺癌细胞MicroRNAs表达的影响.docVIP

  斑蝥素对乳腺癌细胞 MicroRNAs 表达 的影响# 张辉* 5 10 15 20 25 30 35 （上海中医药大学中医复杂系统研究中心，上海 201203） 摘要：目的 观察斑蝥素对乳腺癌细胞 MicroRNAs（miRNAs）表达的影响及分析潜在的治 疗靶点。方法 培养细胞株；MTT 法检测细胞增殖情况；斑蝥素半数抑制浓度（50% inhibiting concentration, IC50）干预 MCF-7 细胞 48h 后，利用 miRNA 芯片技术，检测用药前后 MCF-7 细胞 miRNAs 的表达；采用实时定量 RT-PCR 进行芯片验证和基因表达检测；脂质体法转染 miRNA 抑制物到 MCF-7 细胞；Western blot 检测相关蛋白表达变化。结果 斑蝥素与 MCF-7 细胞生长抑制率之间有浓度依赖关系，48h 的 IC50 为 1.75μg/ml；用 1.75μg/ml 浓度的斑蝥 素干预 MCF-7 细胞 48h 后，与模型组比较，获得 80 个与斑蝥素干预相关 miRNAs（Fold Change≥2，P0.01），其中，上调的 35 个，下调的 45 个；斑蝥素对 miR-106b/93 介导的分 子通路有调控作用。 结论 miRNAs 或许可以作为治疗癌症的药物作用靶标，斑蝥素对 miRNAs 及其介导分子通路的影响可能是其抑制乳腺癌细胞生长的作用途径之一。 关键词：斑蝥素；乳腺癌细胞；miRNAs 中图分类号：R737.9 Cantharidin Alters the Expression of MicroRNAs in Human Breast Cancer Cells ZHANG Hui (Research Center for Traditional Chinese Medicine Complexity System, Shanghai University of Traditional Chinese Medicine, ShangHai 201203) Abstract: Objective To investigate the effects of cantharidin on the microRNA (miRNA) expression in human breast cancer cells and explore the potential targets of the cancer treatment. Methods Cells were grown in RPMI 1640 medium. The inhibition of cell proliferation was measured by MTT assay. MCF-7 cells were treated by cantharidin with 50% inhibiting concentration (IC50) for 48 h. The expression profiles of miRNA in cantharidin-treated and cantharidin-untreated MCF-7 cells were detected with miRNA microarray chips and the array data were verified by quantitative RT-PCR. MCF-7 cells were transiently transfected with miRNA inhibitors by liposome method. Potential mRNA targets were predicted by informatics analysis with TargetScan software. Results Cantharidin significantly inhibited the proliferation of MCF-7 cells in a dose-dependent manner. The IC50 of cantharidin was 1.75μg/ml after treatment for 48 h. The 1.75μg/ml cantharidin was used as the therapeutic drug concentration. Microarray analysis identified 35 miRNAs that were up-regulated and 45 miRNAs that were down-regulated in response to cantharidin tr