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: 10241456
• 负责人: SHUANG HUANG
• 金额: $ 34.29万
• 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/10241456
• 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; Phosphorylation; Play; Protein Isoforms; Protein Kinase; PubMed; Regulation; Reporting; Role; Serous; Signal Pathway; Signal Transduction; Small Interfering RNA; System; Testing; Tissue-Specific Gene Expression; 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; patient derived xenograft model; 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.

项目概要 越来越多的证据清楚地表明 microRNA (miRNA) 系统在 肿瘤相关的上皮间质转化 (EMT) 和肿瘤进展。 研究表明，作为 EMT 抑制因子的 miRNA 远多于促进 EMT 的 miRNA 在卵巢癌中（23 比 3），表明整个 miRNA 系统在卵巢癌中发挥 EMT 抑制作用 这与我们通过沉默 Drosha 和 Dicer 来损害 miRNA 生物发生的观察结果一致。 诱导上皮样卵巢癌细胞出现 EMT 特征，比较 EMT 的丰度。 卵巢癌细胞中的抑制性 miRNA，我们注意到 EMT 抑制性 miRNA 的水平要高得多。 有趣的是，间充质样细胞系中各自原代细胞的水平低于上皮样细胞系。 这些 miRNA 中的大多数 miRNA (pri-miRNA) 在间充质样和上皮样之间是相似的 这些结果表明 miRNA 的生物发生（pri-miRNA 加工成 miRNA）。 在间充质样卵巢癌细胞中效果不佳 我们证明了 miRNA 生物发生的阻断。 需要白细胞介素增强子结合因子 3 (ILF3) 的存在，因为 ILF3 的敲低会增加 为了了解 ILF3 如何抑制 miRNA 生物发生，我们发现了该蛋白质。 激酶 Cδ (PKCδ) 是一种新型 PKC 同工型，可以磷酸化 ILF3，并且 PKCδ 是阻止 ILF3 所必需的 基于这些发现，我们形成了我们的研究。 中心假设：PKCδ 通过赋予 ILF3 促进卵巢癌 EMT 和肿瘤发展 阻止 EMT 抑制性 miRNA 的生物发生的能力这些发现也提供了基础。 开发一种新的卵巢癌靶向治疗方式，即在卵巢癌中建立有效的 miRNA 加工 本申请提出了通过干扰卵巢癌细胞PKCδ功能的三个目标：1) 阐明 PKCδ 调控 miRNA 加工的机制 2) 定义相关机制； PKCδ/ILF3 对卵巢癌细胞 EMT 的调节；以及 3) 研究干扰的潜力； PKCδ抑制卵巢肿瘤发生的功能的成功应用将有助于我们对PKCδ抑制卵巢肿瘤发生的认识。 PKCδ/ILF3 功能轴如何阻断 miRNA 生物发生并展示了抑制的潜力 通过靶向 PKCδ 来抑制卵巢肿瘤发生。