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通过miR-590抑制乳腺癌细胞系MDA-MB-231和MCF-7中参与炎症和癌症的分子信号通路。

Suppressing the molecular signaling pathways involved in inflammation and cancer in breast cancer cell lines MDA-MB-231 and MCF-7 by miR-590.

作者信息

Sheikholeslami Azar, Nabiuni Mohammad, Arefian Ehsan

机构信息

1 Department of Animal Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.

2 Department of Cell and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.

出版信息

Tumour Biol. 2017 Apr;39(4):1010428317697570. doi: 10.1177/1010428317697570.

Abstract

Breast cancer is the most frequent cancer among women worldwide. Tumor immunology suggests relationships between the immune system, chronic inflammation, and cancer. The immune system may either prevent or promote carcinogenesis. Here, we evaluated molecular signaling pathways common in inflammation and cancer and detected the microRNAs which play pivotal roles in mediating these pathways. Using bioinformatics assays, signaling pathways common in inflammation and cancer, and microRNAs mediating these pathways were identified. MiR-590 was selected and cloned into the pLenti-III-eGFP vector and transfected into the breast cancer cell lines. The expression level of microRNA and the candidate genes was evaluated by real-time quantitative reverse transcription polymerase chain reaction, and the apoptosis level in transfected cells was measured by Annexin V-7AAD assay. The cell migration was tested by real-time quantitative reverse transcription polymerase chain reaction for MMP2/MMP9. The expression levels of miR-590 and the selected genes (i.e. JAK2, PI3K, MAPK1, and CREB) were measured 72 h after transfection. While miR-590 showed an over-expression, the genes were significantly down-regulated. A significant increase was observed in apoptosis level in both cell lines and MMP2/MMP9 was significantly decreased in MDA-MB-231 cells. MiR-590 was selected as a microRNA which triggers and down-regulates critical genes of signaling pathways similar in cancer and inflammation. Following the miR-590 treatment, JAK2, PI3K, MAPK1, and CREB were down-regulated and the apoptosis level was increased in breast cancer cell lines. Apparently, some microRNAs can be good candidates for novel treatments of cancer. Although miR-590 showed good results in this study, further studies are required to investigate the role of miR-590 in breast cancer therapy.

摘要

乳腺癌是全球女性中最常见的癌症。肿瘤免疫学揭示了免疫系统、慢性炎症与癌症之间的关系。免疫系统既可能预防癌症发生,也可能促进癌症发生。在此,我们评估了炎症和癌症中常见的分子信号通路,并检测了在介导这些通路中起关键作用的微小RNA。通过生物信息学分析,确定了炎症和癌症中常见的信号通路以及介导这些通路的微小RNA。选择miR-590并克隆到pLenti-III-eGFP载体中,然后转染到乳腺癌细胞系中。通过实时定量逆转录聚合酶链反应评估微小RNA和候选基因的表达水平,并通过膜联蛋白V-7AAD分析检测转染细胞中的凋亡水平。通过实时定量逆转录聚合酶链反应检测MMP2/MMP9来测试细胞迁移。转染72小时后测量miR-590和所选基因(即JAK2、PI3K、MAPK1和CREB)的表达水平。虽然miR-590表现出过表达,但这些基因显著下调。在两种细胞系中均观察到凋亡水平显著增加,并且MDA-MB-231细胞中的MMP2/MMP9显著降低。miR-590被选为一种微小RNA,它触发并下调癌症和炎症中相似的信号通路的关键基因。在miR-590处理后,乳腺癌细胞系中的JAK2、PI3K、MAPK1和CREB下调,凋亡水平增加。显然,一些微小RNA可能是癌症新疗法的良好候选者。尽管miR-590在本研究中显示出良好的结果,但仍需要进一步研究来探究miR-590在乳腺癌治疗中的作用。

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