MOLECULAR PATHOLOGY OF ORAL NEOPLASMS WITH HPV INFECTION

人乳头瘤病毒(HPV)可引起广泛的良性和 在肛门生殖道的恶性疾病，我们已经进行了 全面的分子研究。病毒癌蛋白E6和E7在 它们的结合在启动多步致癌中的主要作用 抑癌蛋白P53与视网膜母细胞瘤易感性 蛋白。利用上皮筏培养，我们研究了细胞因子的功能。 高危型和低危型E6和E7蛋白在分化中的差异 角质形成细胞，并鉴定出新的宿主细胞防御反应， 包括对p21(cip1/waf1/sdi1)和p53的诱导。自.以来 生殖器和口腔粘膜相似，难怪HPV DNA 在头颈部(H&N)的上皮性病变中也检测到 网站。然而，除了喉乳头状瘤之外，没有 人乳头瘤病毒基因在上颌骨中表达的直接分子证据 空气消化系统。我们认为E6和E7基因是活跃的 在H&N肿瘤中转录并促进其发展 港湾人乳头瘤病毒。这项研究旨在提供详细的分子 口腔肿瘤中病毒基因表达和宿主细胞反应的研究 阐述人乳头状瘤病毒在上皮细胞发病中的作用并鉴定 重要的分子预测因子在诊断和治疗中的作用 癌前状态。具体目标是：(1)识别可能的HPV 口腔良性肿瘤和恶性口腔肿瘤的感染 有或没有免疫抑制或致癌风险的新患者 各种因素。病毒类型将通过鉴定PCR产物来确定， 两组DNA靶向粘膜营养型和皮肤型HPV 已在口腔损伤中发现。(2)评估病毒转录和 原位杂交法检测病毒DNA的物理状态 检测、支持或裁定HPV之间的因果关系 和口腔肿瘤。(3)将病毒基因的表达与 不同H&N部位不同级别肿瘤的细胞反应。 P53等细胞周期调控蛋白的可能调控 包括细胞周期蛋白依赖性激酶抑制剂p21、p27KIP1、p57KIP2、 墨水蛋白家族(p15、p16、p18、p19)与转化生长 因子-β1将通过免疫细胞化学(ICC)和 连续切片中的免疫荧光。他们的信使核糖核酸将通过原位杂交进行检测。 以确定调控是转录的还是后转录的 转录的。(4)检测新鲜外科手术中宿主DNA的合成 活组织检查，将可能的非计划染色体复制与 AIM中描述的宿主和病毒基因的表达3.宿主复制 癌前病变和癌前病变中明显分化的超基底细胞 口腔癌将通过掺入3/H-胸腺嘧啶核苷或BrdU进行监测 或通过检测指示DNA复制的细胞蛋白质 能力(例如，增殖细胞核抗原和细胞周期蛋白E)。(5)确定发病时间是否 病毒和宿主基因的表达可能涉及入侵和 转移与肿瘤分级有关。矩阵的生成 金属蛋白酶(MMPs)(如胶原酶1、基质溶酶、基质溶酶1) 和MMPs的组织抑制物(TIMP)1和2将由ICC和 差不多。

Human papillomaviruses (HPVs) induce a wide spectrum of benign and malignant diseases in the anogenital tract, on which we have conducted comprehensive molecular studies. The viral oncoproteins E6 and E7 play a major role in initiating multi-step carcinogenesis through their binding to the tumor suppressor proteins p53 and retinoblastoma susceptibility protein. Using epithelial raft cultures, we investigated the functions of E6 and E7 proteins from high-risk and low-risk genotypes in differentiated keratinocytes and identified novel host cell defensive responses, including the induction of p21 (cip1/waf1/sdi1) and p53. Since the genital and oral mucosa are similar, it is not surprising that HPV DNAs have also been detected in epithelial lesions of head and neck (H&N) sites. Yet, with the exception of laryngeal papillomas, there has been no direct molecular demonstration of HPV gene expression in the upper aerodigestive tract. We propose that the E6 and E7 genes are actively transcribed in and contribute to the development of H&N neoplasms that harbor HPVs. This study is designed to provide a detailed molecular profile of viral gene expression and host cell responses in oral tumors, to elaborate the role of HPVs in epithelial pathogenesis and to identify significant molecular predictors for the diagnosis and treatment of precancerous conditions. Specific aims are: (1) To identify possible HPV infections in benign and malignant oral tumors from archival biopsies and new patients with or without immunosuppressive or carcinogenic risk factors. Virus types will be determined by characterizing PCR products, targeting both mucoso-trophic and cutaneous HPVs since DNAs of both groups have been found in oral lesions. (2) To assess viral transcription and the physical state of the viral DNA by in situ hybridization (ISH) detection, supporting or ruling against causal relationships between HPV and oral neoplasms. (3) To correlate the expression of viral genes and cellular responses in successive grades of neoplasms in various H&N sites. The possible modulation of p53 and other cell cycle regulatory proteins including inhibitors of cyclin-dependent kinases, p21, p27KIP1, p57KIP2, the INK family of proteins (p15, p16, p18, p19), and transforming growth factor-beta1 will be investigated by immunocytochemistry (ICC) and immunofluorescence in serial sections. Their mRNA will be examined by ISH to determine whether regulation is transcriptional or post- transcriptional. (4) To detect host DNA synthesis in fresh surgical biopsies, tieing possible unscheduled chromosomal replication to the expression of host and viral genes described in Aim 3. Host replication in apparently differentiated suprabasal cells in precancerous lesions and oral cancers will be monitored by incorporation of 3/H-thymidine or BrdU or by the detection of cellular proteins indicative of DNA replication capability (e.g., PCNA and cyclin E). (5) To determine whether the onset of expression of viral and host genes potentially involved in invasion and metastasis is linked to tumor grade. The production of matrix metalloproteinases (MMPs) (e.g., collagenase 1, matrilysin, stromolysin 1) and tissue inhibitors of MMPs (TIMP) 1 and 2 will be probed by ICC and ISH.