Cantharidin modulates the E2F1/MCM7-miR-106b-93/p21-PTEN signaling axis in MCF-7 breast cancer cells.
Cantharidin modulates the E2F1/MCM7-miR-106b-93/p21-PTEN signaling axis in MCF-7 breast cancer cells.
复制标题
斑蝥素调节 MCF-7 乳腺癌细胞中的 E2F1/MCM7-miR-106b-93/p21-PTEN 信号轴
DOI：
10.3892/ol.2015.3681
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发表时间：
2015-11
期刊：
影响因子：
2.9
通讯作者：
Yan X
中科院分区：
文献类型：
--
作者：
Zhang H;Yan X
斑蝥素（CTD）是中药中用于治疗癌症的众多天然产物之一。本研究旨在探讨CTD对MCF-7乳腺癌细胞microrna （miRNAs/miRs）表达变化的影响，并探讨其对MCF-7乳腺癌细胞的抗增殖作用。MTT法测定MCF-7细胞的增殖情况。用不同浓度的CTD处理MCF-7细胞48小时，使用miRNA微阵列芯片检测CTD处理和未处理MCF-7细胞中miRNA的表达谱。通过逆转录-定量聚合酶链反应验证阵列数据，western blot检测蛋白表达水平。CTD对MCF-7细胞增殖的50%抑制浓度为1.75µg/ml，且呈剂量依赖性（P<0.01）。此外，微阵列分析发现，35个mirna在CTD治疗中上调（倍数变化≥2.0，P<0.01）， 45个mirna下调（倍数变化≤0.5，P<0.01）。因此，许多ctd诱导的mirna似乎与乳腺癌有关。值得注意的是，CTD抑制了miR-106b-93及其宿主基因MCM7及其转录因子E2F1的表达。此外，CTD诱导miR-106b-93靶基因p21以及磷酸酶和紧张素同源物的蛋白表达水平升高。这些观察结果表明，miRNA表达的调节是CTD在乳腺癌中生物学效应的重要机制。
Cantharidin (CTD) is one of numerous natural products used in traditional Chinese medicine for the treatment of cancer. The aim of the present study was to investigate the effects of CTD on changes in the expression of microRNAs (miRNAs/miRs) and to explore its anti-proliferative effect on MCF-7 breast cancer cells. The proliferation of MCF-7 cells was measured by performing an MTT assay. MCF-7 cells were treated with various concentrations of CTD for 48 h, and the expression profiles of miRNAs in CTD-treated and -untreated MCF-7 cells were detected using miRNA microarray chips. The array data were confirmed by reverse transcription-quantitative polymerase chain reaction and protein expression levels were measured by western blot analysis. The 50% inhibitory concentration of CTD was 1.75 µg/ml following treatment for 48 h and CTD significantly inhibited the proliferation of MCF-7 cells in a dose-dependent manner (P<0.01). Furthermore, microarray analysis identified 35 miRNAs that were up-regulated (fold change ≥2.0 and P<0.01) and 45 miRNAs that were down-regulated (fold change ≤ 0.5 and P<0.01) in response to CTD treatment. Thus, numerous CTD-induced miRNAs appeared to be associated with breast cancer. Notably, CTD repressed the expression of miR-106b-93, its host gene MCM7 and its transcription factor E2F1. In addition, CTD induced an increase in the protein expression levels of miR-106b-93 target genes p21 and phosphatase and tensin homolog. These observations suggested that the modulation of miRNA expression is an important mechanism underlying the biological effects of CTD in breast cancer.