APOPTOSIS INDUCING GENE THERAPY FOR ORAL PREMALIGNANCY

口腔鳞状细胞癌患者 患有恶性疾病，通常表现为晚期疾病，其 管理往往会对这些人的言论产生深远的影响， 吞咽、美容，以及生存。不幸的是， 尽管当代外科手术和放射治疗在过去几年中取得了进展 在过去的30年里，这些患者的存活率基本上保持不变。 原封不动。此外，化疗在治疗中几乎没有什么希望。 这些癌症的管理。因此，开发新的机制， 终止导致这些恶性肿瘤的癌前过程可能会 提供这一医学领域最重大的进步。 口腔癌前病变和口腔原位癌的基因治疗 可能是非常有可能实现的。上皮细胞的局部转导 在癌前过程中经历的转化很可能是 很有效。野生型P53腺病毒的初步调查显示 由于其作为肿瘤的功能，有望成为一种分子干预手段 抑制基因及其在细胞凋亡中的作用。然而，重要的是 需要在理解转导的最优化方面取得进展 分子治疗的效率以及增加我们对 分子干预策略的相互作用以产生 肿瘤细胞死亡的预期效果与非恶性肿瘤的保留 细胞。我们选择使用腺病毒载体作为一种机制 分子干预由于其特有的瞬时表达， 表达效率和扩增到高滴度的能力。 此外，从生物安全的角度来看，缺乏永久融合是有利的 与逆转录病毒载体相比的立场。因此，这项提议 建议进一步加深对凋亡诱导基因的理解 宫颈鳞癌癌前病变和微侵袭的治疗 口腔通过以下具体目标： 1.确定腺病毒载体的最大转染率 标记基因在器官和组织中的瞬时表达 口腔癌前病变的切片模型。 2.检测野生型瞬时表达的有效性 P53、p16、p21(CIP/WAF1)和pRB105在 口腔鳞癌细胞株的诱导凋亡作用及其机制 癌症前疾病的模型。 3.确定转录因子的调控，包括 位于P53下游的P21(CIP/WAF1)、E2F-1和DP-1可以进一步 增强口腔鳞癌细胞的凋亡诱导作用 癌症前的模型。 4.确定优化的基因治疗策略的效果 在癌前口腔癌中诱导细胞凋亡 正常口腔上皮和基质细胞。

Patients with squamous cell carcinoma of the oral cavity are afflicted with a malignancy that frequently presents with advanced disease and whose management often has profound effects upon these individual's speech, swallowing, cosmetic appearance, as well as survival. Unfortunately, despite advances in contemporary surgery and radiation therapy over the past thirty years, survival among these patients has remained essentially unaltered. Additionally, chemotherapy has shown little promise in the management of these cancers. Therefore, developing novel mechanisms of cessating the premalignant processes which lead to these malignancies may provide the most significant advancements in this field of medicine. Gene therapy for premalignancy and carcinoma in-situ of the oral cavity may be very achievable. Topical transduction of epithelium which has undergone transformation in the premalignant process is likely to be efficacious. Initial investigations of wild-type p53 adenovirus has shown promise as a molecular intervention due to its function as a tumor suppressor gene and proposed role in apoptosis. However, significant advances are required in understanding the optimization of transduction efficiency in molecular therapy as well as increasing our understanding of the interaction of molecular intervention strategies in order to produce the desired affect of tumor cell demise and sparing of non-malignant cells. We have elected to employ the adenovirus vector as a mechanism for molecular intervention due to its characteristic transient expression, efficiency of expression, and ability to expand to high titers. Additionally, lack of permanent integration is favorable from a biosafety standpoint as compared to retrovirus vectors. This proposal therefore submits to further develop our understanding of apoptosis inducing gene therapy for premalignant and microinvasive squamous cell carcinoma of the oral cavity through the following specific aims: 1. To determine the maximal transfection efficiency of adenovirus vectors in the transient expression of marker genes in organotypic and tissue slice models of premalignancies of the oral cavity. 2. To determine the effectiveness of transient expression of wild-type p53, p16, p21 (CIP/WAF1), and pRB105 independently and in combination in the induction of apoptosis in oral squamous carcinoma cell lines and the models of premalignancy. 3. To determine whether regulation of transcription factors, including p21 (CIP/WAF1), E2F-1, and DP-1, which are downstream of p53, can further potentiate the induction of apoptosis in oral squamous carcinoma cells and the models of premalignancy. 4. To determine the effect of the optimized gene therapy strategies for induction of apoptosis, in premalignant oral cavity carcinomas, upon normal oral epithelium and stromal cells.