Impact of microRNA processing on EMT of ovarian cancer cells

microRNA加工对卵巢癌细胞EMT的影响

• 批准号: 9768415
• 负责人: SHUANG HUANG
• 金额: $ 33.31万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9768415
• 关键词: Affinity; Binding; Biogenesis; Cancer cell line; Cell Line; Cells; Characteristics; Data; Development; Distant; Down-Regulation; Epithelial; Epithelial Cells; Epithelial ovarian cancer; Greater sac of peritoneum; Growth Factor; Homeobox; Impairment; In Vitro; Knowledge; Malignant Neoplasms; Malignant neoplasm of ovary; Mediating; Mesenchymal; MicroRNAs; Modality; Molecular; Organ; Outcome; Ovary; Patients; Phosphorylation; Play; Protein Isoforms; Protein Kinase; PubMed; Regulation; Reporting; Role; Serous; Signal Pathway; Signal Transduction; Small Interfering RNA; System; Testing; Tissue-Specific Gene Expression; Xenograft Model; anti-cancer therapeutic; aptamer; base; cancer cell; cytokine; inhibitor/antagonist; innovation; knock-down; novel; nuclear factor of activated T-cells, 90 kD; nuclease; ovarian neoplasm; prevent; success; targeted treatment; trait; transcription factor; tumor; tumor microenvironment; tumor progression; tumorigenesis

Project Summary Accumulating evidences have clearly demonstrated that microRNA (miRNA) system plays an active role in tumor-associated epithelial-mesenchymal transition (EMT) and tumor progression. Our PubMed-based search revealed that there are overwhelmingly more miRNAs serving as EMT suppressors than those promoting EMT in ovarian cancer (23 vs 3), indicating that overall miRNA system play an EMT-suppressive role in ovarian cancer. This is consistent with our observation that impairing miRNA biogenesis by silencing Drosha and Dicer induces the occurrence of EMT traits in epithelial-like ovarian cancer cells. Comparing the abundance of EMT- suppressive miRNAs in ovarian cancer cells, we noticed that levels of EMT-suppressive miRNAs are much less in mesenchymal-like cell lines than epithelial-like ones. Intriguingly, levels of the respective primary miRNAs (pri-miRNAs) in majority of these miRNAs are similar between mesenchymal- and epithelial-like ovarian cancer cells. These results indicate that the biogenesis of miRNA (processing of pri-miRNA to miRNA) is not efficient in mesenchymal-like ovarian cancer cells. We showed that blockage of miRNA biogenesis requires the presence of interleukin enhancer-binding factor 3 (ILF3) because knockdown of ILF3 increases miRNA biogenesis. In an effort to understand how ILF3 inhibits miRNA biogenesis, we found that protein kinase Cδ (PKCδ), a novel PKC isoform, can phosphorylate ILF3 and that PKCδ is required for the deterrence of miRNA biogenesis in mesenchymal-like ovarian cancer cells. Based on these findings, we formed our central hypothesis: PKCδ promotes ovarian cancer EMT and tumor development by conferring ILF3 with the ability to deter the biogenesis of EMT-suppressive miRNAs. These findings also provide the basis to develop a novel ovarian cancer-targeted therapeutic modality that is to establish efficient miRNA processing in ovarian cancer cells through the interference of PKCδ function. Three aims are proposed in this application: 1) Elucidate the mechanism underlying PKCδ regulation of miRNA processing; 2) Define mechanisms associated with PKCδ/ILF3 regulation of EMT in ovarian cancer cells; and 3) Investigate the potential of interfering with PKCδ function to suppress ovary tumorigenesis. The success of this application will help our understanding on how PKCδ/ILF3 functional axis blocks miRNA biogenesis and also demonstrate the potential of suppressing ovary tumorigenesis by targeting PKCδ.

项目摘要 积累的证据清楚地表明，microRNA（miRNA）系统在 与肿瘤相关的上皮 - 间质转变（EMT）和肿瘤进展。我们的基于PubMed的搜索 表明，与促进EMT的miRNA相比，有压倒性的miRNA用作EMT补充剂 在卵巢癌（23 vs 3）中，表明整体miRNA系统在卵巢中起抗EMT抑制作用 癌症。这与我们的观察结果一致，即通过沉默的Drosha和Dicer损害miRNA生物发生 诱导上皮样卵巢癌细胞中EMT性状的发生。比较EMT的抽象 我们注意到卵巢癌细胞中的miRNA，我们注意到EMT抑制miRNA水平很大 在间充质样细胞系中，少于上皮样细胞系。有趣的是，相应的主要水平 这些miRNA中的miRNA（pri-miRNA）在间质和上皮样中相似 卵巢癌细胞。这些结果表明miRNA的生物发生（将Pri-miRNA的加工到miRNA） 在间充质样卵巢癌细胞中不有效。我们表明了miRNA生物发生的阻塞 需要白介素增强子结合因子3（ILF3），因为ILF3的敲低增加 miRNA生物发生。为了了解ILF3如何抑制miRNA生物发生，我们发现蛋白 激酶Cδ（PKCδ）是一种新型的PKC同工型，可以磷酸化ILF3，并且确定PKCδ是必需的 间充质样卵巢癌细胞中的miRNA生物发生。根据这些发现，我们成立了 中央假设：PKCδ通过与ILF3与 确定EMT抑制miRNA的生物发生的能力。这些发现也为 开发一种新型的卵巢癌为靶向的治疗方式，是在建立有效的miRNA加工 卵巢癌细胞通过PKCδ功能的干扰。在此应用中提出了三个目标：1） 阐明miRNA加工的PKCδ调节的机制； 2）定义相关的机制 卵巢癌细胞中EMT的PKCδ/ILF3调节； 3）研究干扰的潜力 PKCδ功能可抑制卵巢肿瘤发生。该应用程序的成功将有助于我们理解 PKCδ/ILF3功能轴如何阻止miRNA生物发生，还证明了抑制的潜力 卵巢肿瘤发生通过靶向PKCδ。