Role of MicroRNA 363 in HPV-positive oral cancer

MicroRNA 363 在 HPV 阳性口腔癌中的作用

• 批准号: 8385514
• 负责人: SALEEM A. KHAN
• 金额: $ 21.71万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-12-01 至 2014-03-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8385514
• 关键词: Adhesions; Affect; Alcohols; Apoptosis; Apoptotic; Binding; Cancer cell line; Cell Cycle Regulation; Cell Line; Cell Proliferation; Cell physiology; Cells; Chromatin; Chromatin Structure Alteration; DNA Methylation; Data; Diagnosis; Epigenetic Process; Future; Gene Expression; Gene Expression Regulation; Gene Targeting; Goals; Histone Deacetylase; Histones; Human; Human Papillomavirus; Human papilloma virus infection; Human papillomavirus 16; Incidence; Investigation; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Mediating; Methylation; MicroRNAs; Molecular; Molecular Profiling; Mouth Carcinoma; Mutation; Nature; Normal tissue morphology; Oncogene Proteins; Oncogenes; Oncogenic; Oral; Oral mucous membrane structure; Oropharyngeal Squamous Cell Carcinoma; Pathogenesis; Pathway interactions; Patients; Process; Proteins; Publishing; Regulation; Regulator Genes; Role; Small Interfering RNA; Squamous cell carcinoma; Testing; Tissues; Tobacco use; Transferase; Tumor Suppressor Proteins; Viral; Viral Cancer; Virus; base; carcinogenesis; cell growth; histone methyltransferase; histone modification; inhibitor/antagonist; keratinocyte; knock-down; malignant mouth neoplasm; malignant oropharynx neoplasm; migration; mortality; new therapeutic target; oral carcinogenesis; outcome forecast; overexpression; promoter; research study; response; therapeutic target; tobacco exposure; transcription factor; tumor progression

DESCRIPTION (provided by applicant): Recently, there has been a substantial increase in the incidence of human papillomavirus (HPV)-associated oropharyngeal (OP) squamous cell carcinoma (OPSCC), while other cases result from alcohol and/or tobacco use. Since HPV infection confers a significantly decreased mortality rate, understanding the molecular mechanisms responsible for HPV-associated oral carcinoma is crucial to understanding the pathways regulated by HPV versus those involved in HPV-negative oral cancer. Recently, a subpopulation of HPV-positive OPSCC patients with tobacco exposure and p53 genetic alterations has been identified, warranting detailed investigation of the HPV E6- and p53-mediated pathways contributing to carcinogenesis. In HPV-associated OPSCC, the viral E6 and E7 oncoproteins are expressed at high levels. MicroRNAs (miRNAs) are ~22 nt long single-stranded RNAs that generally function as negative regulators of gene expression, and their expression profiles are altered in a variety of human cancers. Limited information is available on the role of miRNAs in the pathogenesis of OP cancers. Our published studies using OPSCC cell lines show that miRNA expression profiles in HPV-positive and HPV-negative OP cancers are distinctively different from each other, including the overexpression of miR-363 and underexpression of miR-218 in HPV-positive cell lines. MiR-363 is also overexpressed in HPV-16 positive OP cancer tissues compared to HPV-negative OPSCC tissues, normal oral mucosa and normal oral keratinocytes (NOKs). Our data also show that the HPV-16 E6 oncogene promotes miR-363 overexpression while reducing miR-218 expression. We hypothesize that differences in miR-363 and/or miR-218 levels change the expression of their target genes and this alters the cellular pathways involved in HPV-positive oral carcinogenesis. In Aim 1, we will test whether miR-363 and miR- 218 are oncogenic and tumor suppressor miRNAs, respectively, that affect important cellular pathways. We will accomplish this by overexpressing or knocking-down the expression of miR-363 and miR-218 in HPV-positive and HPV-negative OPSCC cell lines, NOKs and NOK expressing E6, and study their effect on cell proliferation, migration, invasion, adhesion and apoptosis. In Aim 2, we will identify the regulatory mechanisms by which E6 affects miR-363 and miR-218 expression and whether these are p53-dependent. We will use inhibitors of DNA methylation and histone modification to test whether epigenetic processes are involved in E6-mediated regulation of these miRNAs. If this is the case, we will also study the methylation status and chromatin state of these miRNA promoters. We will also test whether E6 affects miR-363 and miR-218 expression through inactivation of the p53 pathway. Our studies should provide valuable information on the role of miR-363 and miR-218 in HPV-positive OP cancers, the nature of regulation of these miRNAs by E6, and the molecular basis for the differential regulation of miR-363 and miR-218 in HPV-positive and HPV-negative OPSCC. This information could be utilized for future diagnosis, prognosis and therapeutic targets for these cancers.

描述（由申请人提供）：最近，人乳头瘤病毒（HPV）相关口咽（OP）鳞状细胞癌（OPSCC）的发病率大幅增加，而其他病例则由饮酒和/或吸烟引起。由于 HPV 感染可显着降低死亡率，因此了解 HPV 相关口腔癌的分子机制对于了解 HPV 调节的途径与 HPV 阴性口腔癌中涉及的途径至关重要。最近，已鉴定出存在烟草暴露和 p53 基因改变的 HPV 阳性 OPSCC 患者亚群，需要对 HPV E6 和 p53 介导的致癌途径进行详细研究。在 HPV 相关的 OPSCC 中，病毒 E6 和 E7 癌蛋白高水平表达。 MicroRNA (miRNA) 是约 22 nt 长的单链 RNA，通常充当基因表达的负调节因子，并且它们的表达谱在多种人类癌症中发生改变。关于 miRNA 在 OP 癌症发病机制中的作用的信息有限。我们发表的使用 OPSCC 细胞系的研究表明，HPV 阳性和 HPV 阴性 OP 癌症中的 miRNA 表达谱彼此明显不同，包括 HPV 阳性细胞系中 miR-363 的过度表达和 miR-218 的表达不足。与 HPV 阴性 OPSCC 组织、正常口腔粘膜和正常口腔角质形成细胞 (NOK) 相比，miR-363 在 HPV-16 阳性 OP 癌组织中也过表达。我们的数据还表明，HPV-16 E6 癌基因促进 miR-363 过度表达，同时减少 miR-218 表达。我们假设 miR-363 和/或 miR-218 水平的差异改变了其靶基因的表达，从而改变了与 HPV 阳性口腔癌发生相关的细胞途径。在目标 1 中，我们将测试 miR-363 和 miR-218 是否分别是影响重要细胞通路的致癌和抑癌 miRNA。我们将通过在HPV阳性和HPV阴性OPSCC细胞系、NOK和表达E6的NOK中过表达或敲低miR-363和miR-218的表达来实现这一目标，并研究它们对细胞增殖、迁移、侵袭、粘附和凋亡的影响。在目标 2 中，我们将确定 E6 影响 miR-363 和 miR-218 表达的调节机制以及这些机制是否依赖于 p53。我们将使用 DNA 甲基化和组蛋白修饰的抑制剂来测试表观遗传过程是否参与 E6 介导的这些 miRNA 的调节。如果是这样的话，我们还将研究这些miRNA启动子的甲基化状态和染色质状态。我们还将测试 E6 是否通过 p53 通路失活来影响 miR-363 和 miR-218 的表达。我们的研究应该提供有价值的信息，包括 miR-363 和 miR-218 在 HPV 阳性 OP 癌症中的作用、E6 对这些 miRNA 的调节性质，以及 miR-363 和 miR-218 在 HPV 阳性和 HPV 阴性 OPSCC 中差异调节的分子基础。这些信息可用于这些癌症的未来诊断、预后和治疗目标。

项目成果

MicroRNA-363 targets myosin 1B to reduce cellular migration in head and neck cancer.

• DOI: 10.1186/s12885-015-1888-3
• 发表时间: 2015-11-06
• 期刊: BMC cancer
• 影响因子: 3.8
• 作者: Chapman BV;Wald AI;Akhtar P;Munko AC;Xu J;Gibson SP;Grandis JR;Ferris RL;Khan SA
• 通讯作者: Khan SA

Identification of miRNAs dysregulated in human foreskin keratinocytes (HFKs) expressing the human papillomavirus (HPV) Type 16 E6 and E7 oncoproteins.

鉴定表达人乳头瘤病毒 (HPV) 16 型 E6 和 E7 癌蛋白的人包皮角质形成细胞 (HFK) 中失调的 miRNA。

• DOI: 10.2174/2211536611302010002
• 发表时间: 2013
• 期刊: MicroRNA (Shariqah, United Arab Emirates)
• 作者: Yablonska,Svitlana;Hoskins,ElizabethE;Wells,SusanneI;Khan,SaleemA
• 通讯作者: Khan,SaleemA