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.

项目摘要 越来越多的证据清楚地表明，microRNA(MiRNA)系统在 肿瘤相关上皮间充质转化(EMT)与肿瘤进展我们基于PubMed的搜索 研究发现，作为EMT抑制因子的miRNAs远远多于那些促进EMT的miRNAs 在卵巢癌中(23对3)，表明整个miRNA系统在卵巢中发挥EMT抑制作用 癌症。这与我们的观察结果一致，即通过沉默DROSHA和DICER来损害miRNA的生物发生 诱导上皮样卵巢癌细胞出现EMT特征。比较EMT的丰度- 在卵巢癌细胞中抑制miRNAs，我们注意到抑制EMT的miRNAs的水平很高 间充质样细胞系比上皮样细胞系少。耐人寻味的是，各个初选的水平 其中大部分miRNAs(pri-miRNAs)在间充质样和上皮样miRNAs中相似 卵巢癌细胞。这些结果表明，miRNA的生物发生(将pri-miRNA加工成miRNA) 对间充质样卵巢癌细胞无效。我们证明了对miRNA生物发生的阻断 需要存在白细胞介素增强子结合因子3(ILF3)，因为ILF3的敲除增加 MiRNA生物发生。为了了解ILF3如何抑制miRNA的生物发生，我们发现 激酶Cδ(PKCδ)是一种新的PKC亚型，它能磷酸化ILF3，而PKCδ是其发挥威慑作用所必需的 间充质样卵巢癌细胞中miRNA生物发生的研究。基于这些发现，我们形成了我们的 中心假说：PKCδ通过激活ILF3促进卵巢癌转移和肿瘤发展 阻止EMT抑制miRNAs生物发生的能力。这些发现也为 开发一种新的卵巢癌靶向治疗方法，即在 卵巢癌细胞通过干扰蛋白激酶C的δ功能发挥作用。本申请提出三个目标：1) 阐明PKCδ调控miRNA加工的潜在机制；2)定义相关机制 用PKCδ/ILF3调控卵巢癌细胞的EMT；3)研究干扰卵巢癌细胞EMT的可能性 PKCδ具有抑制卵巢肿瘤发生的作用。这项申请的成功将有助于我们对 PKCδ/ILF3功能轴如何阻止miRNA的生物发生并证明其抑制的可能性 靶向PKCδ的卵巢肿瘤发生。