BIOCHEMICAL AND CYTOGENEITC MARKERS IN RETINOBLASTOMA

本提案的长期目标是了解 视网膜母细胞瘤（RB）基因，即RB功能的丧失， 人类肿瘤的发展及其作为预后指标的潜力 肿瘤患者的临床结局指标，其中 发生RB功能丧失。 初步研究将集中在 如肺癌、乳腺癌、膀胱癌、骨癌、卵巢癌和 肾脏以及急性粒细胞白血病。 尽可能正常 将检查来自同一部位的组织以及转移病灶 从同一个病人身上取下 主要重点将放在检查RB状态的人类肿瘤 在冷冻和石蜡切片中的单细胞水平， 直接Western免疫印迹，虽然许多肿瘤将 在RNA和DNA水平上进行检测。 后一项将完成 在我们试图找到的结构变化， RB基因导致功能丧失， 蛋白质研究 石蜡切片的使用将在 因为我们最近成功地获得了优秀的 石蜡切片的免疫组化RB染色， 多克隆RB抗体，据我们所知， 任何可商购的RB抗体，无论是多克隆的还是 单克隆的。 实现这一建议的目标， 更好地了解RB基因在一些最重要的疾病中的作用， 人类常见的恶性肿瘤， 确定是否监测特定患者的RB状态 恶性肿瘤将提供与其 临床结果或其疾病的侵袭性。

The long term objective of this proposal is to understand the role of the retinoblastoma (RB) gene, namely loss of RB function, in the development of human tumors as well as its potential to be a prognostic indicator of the clinical outcome in patients with tumors in which the loss of RB function has occurred. Initial studies will concentrate on such tumors as cancers of the lung, breast, bladder, bone, ovary and kidney as well as acute myeloblastic leukemia. Whenever possible normal tissue from the same site will be examined as will metastatic lesions from the same patient when available. Major emphasis will be placed on examining the RB status of human tumors at the single cell level in frozen and paraffin sections as well as direct Western immunoblotting, although many of the tumors will be examined in addition at the RNA and DNA levels. The latter will be done in our attempt to find the structural changes that have occurred within the RB gene to cause the loss of function and to confirm and complement our protein studies. The use of paraffin sections will be done whenever possible since we have recently been successful in obtaining excellent immunohistochemical RB staining of paraffin sections using our polyclonal RB antibody, which, to our knowledge, is not possible with any commercially available RB antibody, whether polyclonal or monoclonal. The fulfillment of the goals of this proposal should provide a much better understanding of the role of the RB gene in some of the most common human malignancies and equally or perhaps even more importantly determine whether monitoring the RB status in patients with specific malignancies will provide significant information related to their clinical outcome or the aggressiveness of their disease.