朱克鹏, 肖勋蓉, 罗义, 弋文, 尹均明, 杨川, 杜果城. miRNA-613在乳腺癌中的表达及作用机制[J]. 中国肿瘤临床, 2019, 46(14): 718-722. DOI: 10.3969/j.issn.1000-8179.2019.14.676
引用本文: 朱克鹏, 肖勋蓉, 罗义, 弋文, 尹均明, 杨川, 杜果城. miRNA-613在乳腺癌中的表达及作用机制[J]. 中国肿瘤临床, 2019, 46(14): 718-722. DOI: 10.3969/j.issn.1000-8179.2019.14.676
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Zhu Kepeng, Xiao Xunrong, Luo Yi, Yi Wen, Yin Junming, Yang Chuan, Du Guocheng. Expression and mechanism of microRNA-613 in breast cancer[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2019, 46(14): 718-722. DOI: 10.3969/j.issn.1000-8179.2019.14.676
Citation: Zhu Kepeng, Xiao Xunrong, Luo Yi, Yi Wen, Yin Junming, Yang Chuan, Du Guocheng. Expression and mechanism of microRNA-613 in breast cancer[J]. CHINESE JOURNAL OF CLINICAL ONCOLOGY, 2019, 46(14): 718-722. DOI: 10.3969/j.issn.1000-8179.2019.14.676
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miRNA-613在乳腺癌中的表达及作用机制

Expression and mechanism of microRNA-613 in breast cancer

    摘要
  • 摘要:
      目的  探讨乳腺癌中微小RNA (microRNA，miRNA)-613表达及作用机制。
      方法  收集2017年5月至2018年5月91例于南充市中心医院手术切除的乳腺癌患者的组织标本，实时荧光定量PCR检测乳腺癌组织及癌旁组织标本、乳腺癌细胞系(MDAMB-231、MDA-MB-468、MCF-7)和正常乳腺上皮细胞系HBL-100中miRNA-613的表达水平，分析其与乳腺癌患者临床病理特征的关系。TCGA数据库分析miRNA-613与乳腺癌患者预后的关系。双荧光素酶报告实验检测miRNA-613与SOX9的3'UTR区的结合情况。将miRNA-613模拟物转染至MDA-MB-231细胞，CCK-8法和Transwell侵袭及迁移实验分别检测细胞增殖活性、侵袭和迁移能力的变化，Western blot检测细胞中SOX9、β-catenin、E-Cadherin和Vimentin蛋白的表达变化。
      结果  miRNA-613在乳腺癌组织中表达明显低于癌旁组织(P < 0.05)，并且miRNA-613表达与TNM分期和淋巴结转移密切相关(P < 0.05)，TCGA生存数据显示miRNA-613表达与乳腺癌患者的总生存率无关(P>0.05)。乳腺癌细胞系中miRNA-613的表达明显低于正常乳腺上皮细胞系(P < 0.05)，并且高侵袭转移性乳腺癌细胞系MDA-MB-231、MDA-MB-468中miRNA-613的表达明显低于低侵袭转移性乳腺癌细胞系MCF-7(P < 0.05)。双荧光素酶报告实验显示miRNA-613可与SOX9的3'UTR特异性结合。上调miRNA-613的表达能抑制MDA-MB-231细胞的增殖和侵袭迁移能力(P < 0.05)，同时下调SOX9、β-catenin和Vimentin蛋白的表达(P < 0.05)，并上调ECadherin蛋白的表达(P < 0.05)。
      结论  在乳腺癌组织和细胞中miRNA-613异常低表达，miRNA-613可能通过调控SOX9、Wnt/β-catenin信号通路抑制乳腺癌细胞的增殖、侵袭转移及上皮间质转化。

     

    Abstract:
      Objective  To investigate the expression and mechanism of microRNA (miRNA)-613 in breast cancer.
      Methods  A total of 91 specimens of breast cancer tissue were collected from Nanchong Central Hospital between May 2017 and May 2018. Quantitative realtime PCR (qRT-PCR) was used to estimate miRNA-613 expression levels in breast cancer and adjacent tissues and breast cancer (cells MDAMB-231, MDA-MB-468, and MCF-7) and normal breast epithelial (HBL-100) cell lines. Based on these data, the relationship between miRNA-613 expression and clinicopathological characteristics and prognosis in breast cancer patients were analyzed using the cancer genome atlas (TCGA) database. A dual-luciferase reporter assay was used to detect the binding of miRNA-613 to the 3'UTR of SOX9. Effects on cell proliferation and cell invasion and migration upon transfection of MDA-MB-231 cells with miRNA-613 mimics were detected by the CCK-8 assay and Transwell invasion and migration assays, respectively. In addition, Western blot was used to estimate the expression levels of SOX9, β-catenin, E-cadherin, and Vimentin in the transfected cells.
      Results  The expression of miRNA-613 in breast cancer tissues was significantly lower than that in adjacent tissues (P < 0.05) and was found to be closely related to TNM stage and lymph node metastasis (P < 0.05). TCGA survival data showed that miRNA-613 expression was not related to the overall survival rate of breast cancer patients (P>0.05). The expression of miRNA-613 in breast cancer cell lines was significantly lower than that in the normal breast epithelial cell line (P < 0.05). Similarly, the expression of miRNA-613 in highly invasive metastatic breast cancer cell lines (MDA-MB-231 and MDA-MB-468) was significantly lower than that in the metastatic breast cancer cell line MCF-7 with lower invasion ability (P < 0.05). The dual-luciferase reporter assay showed that miRNA-613 could specifically bind to the 3'UTR of SOX9. Upregulation of miRNA-613 expression could inhibit the proliferation, invasion, and migration of MDA-MB-231 cells (P < 0.05). This was associated with the downregulated expression of SOX9, β-catenin, and Vimentin (P < 0.05) and upregulation of E-Cadherin expression (P < 0.05).
      Conclusions  The expression of miRNA-613 was decreased in breast cancer tissues and cell lines. MiRNA-613 may inhibit the proliferation, invasion, and metastasis of breast cancer cells and epithelial-mesenchymal transition by regulating the SOX9 and Wnt/β-catenin signaling pathways.

     

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