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，人类乳头瘤病毒（HPV）相关的口咽（OP）鳞状细胞癌（OPSCC）的发生率大幅增加，而其他情况则是由于酒精和/或烟草的使用而引起的。由于HPV感染赋予了死亡率显着降低，因此了解导致与HPV相关的口腔癌的分子机制对于理解由HPV调节的途径与参与HPV阴性口腔癌的途径至关重要。最近，已经确定了烟草暴露和p53遗传改变的HPV阳性OPSCC患​​者的亚群，保证对HPV E6-和p53介导的途径进行详细研究，导致癌变。在与HPV相关的OPSCC中，病毒E6和E7癌蛋白在高水平上表达。 microRNA（miRNA）是〜22 nt长的单链RNA，通常用作基因表达的负调节剂，并且在各种人类癌症中其表达谱发生了变化。关于miRNA在OP癌症发病机理中的作用的信息有限。我们使用OPSCC细胞系发表的研究表明，HPV阳性和HPV阴性OP癌症中的miRNA表达谱彼此截然不同，包括miR-363的过表达以及在HPV阳性细胞系中miR-218的过表达。与HPV阴性OPSCC组织，正常口服粘膜和正常的口腔角质形成细胞（NOKS）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致癌基因促进了miR-363过表达，同时降低了miR-218的表达。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，这改变了与HPV阳性口服癌变有关的细胞途径。在AIM 1中，我们将测试miR-363和miR-218是否分别是影响重要细胞途径的致癌和肿瘤抑制miRNA。我们将通过过度表达或敲击MiR-363和miR-218在HPV阳性和HPV阴性OPSCC细胞系，NOKS和NOK表达E6中的表达来实现这一目标，并研究其对细胞增殖，迁移，迁移，入侵，粘附，粘附，粘附和凋亡的影响。在AIM 2中，我们将确定E6影响miR-363和miR-218表达的调节机制，以及它们是否依赖于p53。我们将使用DNA甲基化和组蛋白修饰的抑制剂测试表观遗传过程是否参与了这些miRNA的E6介导的调节。如果是这种情况，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途径失活而影响miR-363和miR-218。我们的研究应提供有关miR-363和miR-218在HPV阳性OP癌中的作用，E6调节这些miRNA的性质以及miR-363和miR-218在HPV阳性和HPV-Negation-negative opscc中对MiR-363和miR-218的差异调节的分子基础的有价值的信息。这些信息可以用于这些癌症的将来的诊断，预后和治疗靶标。

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