METALLOTHIONEIN AND CYTOTOXIC DRUG RESISTANCE

该奖项的目的是发展一个医生科学家， 癌症药理学 候选人表现出了 有助于临床泌尿肿瘤学，并致力于学术 事业 然而，他缺乏正规的基础科学训练。 赞助商是一个 公认的癌症研究人员，并有培训记录 在基础科学环境中的医生。 拟议的调查 金属硫蛋白（MT）和细胞毒性药物耐药性的研究旨在培养 临床医生在分子生物学，药理学， 实验疗法，并确定新的战略，以克服肿瘤 对化疗的抵抗力。 该研究提案旨在调查MT在以下方面的作用： 赋予对顺铂（cDDP）和相关抗癌剂的抗性。 cDDP对多种人类肿瘤具有活性，据信其作用方式是 在染色体DNA中形成共价交联。 可惜 肿瘤细胞产生的抗性限制了治疗的有效性 cDDP的。 MT是一种富含半胱氨酸的蛋白质，可保护细胞免受重金属的侵害。 金属毒性，是调节细胞反应的重要候选物， 细胞毒性药物，特别是与巯基反应的药物。 我们将 利用酶检测耐药细胞系中MT的升高 使用针对MT的抗体的连接免疫吸附测定（ELISA）。 以来 人MT以几种异构体形式存在，单克隆抗体特异性 将开发每种人类同种型的抗体，并使用ELISA检测 如果耐药细胞系表现出任何特定的 山 此外，将在以下细胞中测定对cDDP的敏感性： MT水平由MT的既定诱导剂操纵。 过的机制 MT的表达将通过表征MT基因拷贝数来研究， MT的mRNA水平和信息的稳定性。 最后，mRNA和蛋白质 MT水平将在来自患有以下疾病的患者的组织样品中测定： 泌尿系恶性肿瘤，以评价MT是否影响对cDDP的反应性， cDDP是否改变MT的人肿瘤表达。 匹兹堡大学特别适合 发展肿瘤学的医生研究者，因为它 匹兹堡癌症研究所最近被授予癌症中心 从NIH获得三年的资助。

The purpose of this award is the development of a physician scientist in cancer pharmacology. The candidate has demonstrated a potential to contribute to clinical urologic oncology and is committed to an academic career. However, he lacks formal basic science training. The sponsor is a recognized cancer researcher with an established record for training physicians in a basic science environment. The proposed investigation of metallothionein (MT) and cytotoxic drug resistance seeks to train a clinician in the discipline of molecular biology, pharmacology, and experimental therapeutics and to identify new strategies to overcome tumor resistance to chemotherapy. The research proposal is designed to investigate the role of MT in conferring resistance to cis-platin (cDDP) and related anti-cancer agents. cDDP is active against a variety of human tumors and is believed to act by forming covalent cross links in chromosomal DNA. Unfortunately, the development of resistance by tumor cells limits the therapeutic usefulness of cDDP. MT, a cysteine rich protein that protects cells against heavy metal toxicity, is an important candidate to modulate cellular response to cytotoxic drugs, especially those reactive with sulfhydryl groups. We will examine the elevation of MT in drug resistant cell lines utilizing enzyme linked immunosorbent assay (ELISA) using antibodies against MT. Since human MT exists in several isomeric forms, monoclonal antibodies specific for each human isoform will be developed and ELISA will be used to examine if drug-resistant cell lines exhibit increases in any particular isoform of MT. In addition, the sensitivity to cDDP will be determined in cells with MT levels manipulated by established inducers of MT. The mechanism of over expression of MT will be studied by characterizing the MT gene copy number, mRNA level and stability of message of MT. Finally, mRNA and protein levels of MT will be determined in tissue samples from patients with urologic malignancy to evaluate if MT influences responsiveness to cDDP and whether cDDP alters human tumor expression of MT. The University of Pittsburgh is particularly well suited for the development of physician investigators in oncology because of its Pittsburgh Cancer Institute which was recently awarded a Cancer Center Support Grant for three years from the NIH.