RAF ONCOGENE ANALYSIS AND RADIATION RESISTANT TUMOR CELL

寻找基因变化的原因和影响 在癌症研究中占有中心地位。 一年半前，我们开始着手解决 一种人类喉癌 我们现在已经报道了 异常RAF癌基因与放射抵抗人类 喉癌细胞系SQ-20 B。 我们的研究 在另一份报告中提出的独立意见作了补充， 非癌性抗辐射皮肤成纤维细胞实验室 有耐辐射家族史的患者 恶性肿瘤显示RAF癌基因异常。 但这些 两份报告尚未证明这一点的确切性质， 异常，也不与靶细胞对 辐射毒性 我们在本赠款期内的主要目标将是 研究RAF基因表达的调节和细胞凋亡的影响。 对辐射损害的反应， 将正义或反义RAF最佳表达至适当的 哺乳动物细胞（AIM I），解剖和表征RAF SQ-20 B细胞（AIM II）中的癌基因结构和功能， 在另外三个密切相关的基因异常中寻找 相关人类癌症（AIM III）。 我们将使用一种明确定义的、可行的和综合的方法， 评估SQ中RAF故障的遗传基础- 20 B细胞。 这些研究极大地促进了可用性 克隆和确定的独特表达载体， RAF（正义/反义）序列，c-RAF-1 DNA探针，核糖核酸探针 适合RAF（反义+）分析的载体探针，和 来自Dr.乔治Mark（NIH）的RAF蛋白抗体。 我们将 继续与阿纳托利博士合作 德里奇洛、乔治马克和迈克尔·戈特斯曼， 放射生物学、分子生物学、 和多药耐药性的分子生物学， 分别 最后，目前的调查应使我们 开始阐明RAF原蛋白的结构和功能， 对维持正常细胞至关重要的癌基因结构域 扩散和行为。

The search for the cause and effect of genetic changes associated with neoplastic cells occupies a central place in cancer research. A year and a half ago, we began to address the possible causes of a human laryngeal cancer. We have now reported the association of an abnormal RAF oncogene with a radiation resistant human laryngeal carcinoma cell line, SQ-20B. Our studies have been complemented by the independent observations made in another laboratory that noncancerous, radiation resistant skin fibroblasts of patients with a familial history of radiation resistant malignancies reveal an abnormal RAF oncogene. However, these two reports have not yet proven the precise nature of this abnormality, nor its relevance to the resistance of target cells to radiation toxicity. Our major gaols during the tenure of this grant period will be to study the modulation of RAF gene expression and the cellular responses to radiation-damage as a consequence of delivering an optimal expression of sense or anti-sense RAF to the appropriate mammalian cells (AIM I), to dissect and characterize the RAF oncogene structure and function in SQ-20B cells (AIM II), and to search for genetic abnormalities in three additional closely related human cancers (AIM III). We will use a clearly defined, feasible, and integrated approach to evaluate the genetic basis of malfunctioning of the RAF in SQ- 20B cells. These studies are greatly facilitated by the availability of cloned and defined unique expression vectors containing human RAF(sense/anti-sense) sequence, c-RAF-1 DNA probes, riboprobe vector probes suitable for RAF (anti-sense+) analysis, and antibody to RAF protein from Dr. George Mark (NIH). We will maintain our ongoing collaborative efforts with Drs. Anatoly Dritschilo, George Mark, and Michael Gottesman for their specific expertise in the areas of radiobiology, molecular biology of RAF, and molecular biology of multidrug resistance, respectively. Finally, the current investigations should lead us to begin to elucidate the structure and function of RAF proto- oncogene domains pivotal to the maintenance of normal cell proliferation and behavior.