Department of Biotechnology, Indian Institute of Technology Madras, Chennai, India.
Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel.
Mol Cell Biol. 2019 Mar 19;39(7). doi: 10.1128/MCB.00577-18. Print 2019 Apr 1.
Breast cancer is a recurrent type of cancer among women worldwide. Despite remarkable progress in the prevention, detection, and treatment of breast cancer, it still remains a major chronic problem worldwide and poses significant challenges, like metastasis to distant organs, demanding the need for novel biomarkers and therapeutic targets. Focal adhesion kinase (FAK), a member of the protein tyrosine kinases, has been shown to be expressed in high levels in breast tumors. Of late, FAK has emerged as an impending curative target in breast carcinoma, with few of the small molecular inhibitors reaching the clinical trial stage. In the current study, we established that microRNA 551a (miR-551a) precisely regulates FAK by binding to the complementary sequences in the 3' untranslated region (UTR) of mRNAs of FAK and inhibits its expression in breast carcinoma cell lines. Further, results from human breast carcinoma samples illustrated that miR-551a levels were substantially downregulated in tumor samples, with a concurrent rise in the expression of FAK. Functional experimental studies using miR-551a-overexpressing breast cancer cells and nude mouse xenograft models revealed the tumor suppressor role of miR-551a. We also found that miR-551a expression decreased the invasion and migratory ability of breast carcinoma cells by inhibiting MMP-9 activity. Regulation studies performed utilizing promoter luciferase assays, chromatin immunoprecipitation (ChIP), and electrophoretic mobility shift assay (EMSA) revealed that c-Fos binds to the miR-551a promoter and activates it. Further, we observed a considerable increase in the amount of miR-551a levels upon c-Fos overexpression. All of these results showed that miR-551a can be of clinical relevance in understanding the regulation of FAK in breast tumorigenesis.
乳腺癌是全球女性中常见的复发性癌症。尽管在预防、检测和治疗乳腺癌方面取得了显著进展,但它仍然是全球的一个主要慢性问题,并带来了重大挑战,如转移到远处器官,这需要新的生物标志物和治疗靶点。粘着斑激酶(FAK)是一种蛋白质酪氨酸激酶,已被证明在乳腺癌肿瘤中高表达。最近,FAK 已成为乳腺癌的一个有前途的治疗靶点,少数小分子抑制剂已进入临床试验阶段。在本研究中,我们发现 microRNA 551a(miR-551a)通过与 FAK mRNA 的 3'非翻译区(UTR)中的互补序列结合,精确调节 FAK,并抑制其在乳腺癌细胞系中的表达。进一步,人类乳腺癌样本的结果表明,miR-551a 在肿瘤样本中的水平显著下调,同时 FAK 的表达升高。使用 miR-551a 过表达的乳腺癌细胞和裸鼠异种移植模型进行的功能实验研究揭示了 miR-551a 的肿瘤抑制作用。我们还发现,miR-551a 通过抑制 MMP-9 活性降低了乳腺癌细胞的侵袭和迁移能力。利用启动子荧光素酶报告基因检测、染色质免疫沉淀(ChIP)和电泳迁移率变动分析(EMSA)进行的调控研究表明,c-Fos 结合到 miR-551a 启动子并激活它。此外,我们观察到 c-Fos 过表达后 miR-551a 水平显著增加。所有这些结果表明,miR-551a 在理解 FAK 在乳腺癌发生中的调节方面具有临床相关性。