GENE EXPRESSION INDICES ASSOCIATED WITH CHEMORESISTANCE

描述：(申请人摘要)非小细胞肺癌 对目前的化疗方案反应差，总体上 退化率只有30-50%。抗药性的机制可能会 涉及多种基因产物。在初步研究中，H1435非小细胞 肺癌(NSCLC)细胞株对卡铂的耐药性是 H460。20个可能与化疗耐药相关的基因的评估 发现谷胱甘肽转移酶(GST)P1、Baxα、GADD45、ERCC3和 谷胱甘肽过氧化物酶、MGST基因在第100、20、10、6、5和 H1435的水平分别高出4倍。主要的长期目标是 建议的调查是为了提高对非小细胞肺癌的机械性理解。 化疗耐药性和开发一种预测哪些非小细胞肺癌肿瘤将 对卡铂有反应。实现这些目标将具有特定的价值1) 允许基于知识开发改进的化疗药物 机制，2)挽救卡铂耐药肿瘤患者 避免无效治疗带来的不必要的痛苦和费用，3) 允许卡铂耐药肿瘤患者寻求替代方案 不拖延的治疗选择，以及4)提供一个通用模型 识别对其他化疗方案耐药的肿瘤。要实现 这些目标，很多基因都必须进行评估。癌细胞株将成为 最适合进行全面调查的模式。具体目标是 以下是。目的1)确定80个推定的卡铂的表达 检测10株非小细胞肺癌耐药基因的IC30浓度 卡铂用于每一行，然后确定哪一种构成表达 模式在统计上的相关性最好(通过线性回归或邓肯的 多范围测试)，带有电阻。任何确定的关联都将是 在另外十条非小细胞肺癌线路上进行了验证。目标2)结合个人 将构成的基因表达值转化为指数并对其进行评估 与最初的10个耐药细胞株相关联。任何关联 将在第二个十行中进行验证。竞争性模板RT-PCR基因 为本项目提出的表情量化方法是由 申请者是最好的，因为1)它允许高吞吐量 对许多样品进行测量。2)所产生的数据采用标准单位，以便 单独的基因表达值可以在数学上组合在基因中 表达指数，以及3)因为相同的竞争性标准混合物 模板将用于所有拟议的实验，数据来自所有 可以对实验进行比较以进行统计分析。

DESCRIPTION: (Applicant's Abstract) Non-small cell lung cancer (NSCLC) is poorly responsive to current chemotherapeutic regimens with an overall regression rate of only 30-50%. The mechanisms of resistance are likely to involve multiple gene products. In preliminary studies the H1435 non-small cell lung cancer (NSCLC) cell line is 50-fold more resistant to carboplatin than H460. Evaluation of 20 genes putatively associated with chemoresistance revealed that glutathione transferase (GST) P1, Bax alpha, GADD45, ERCC3, and glutathione peroxidase, mGST genes are expressed at 100, 20, 10, 6, 5, and 4-fold higher levels respectively in H1435. The primary long term objective of the proposed investigation is to improve mechanistic understanding of NSCLC chemoresistance and to develop a method for predicting which NSCLC tumors will respond to carboplatin. Achieving these goals will have specific value by 1) allowing development of improved chemotherapeutic agents based on a knowledge of mechanisms, 2) sparing patients with carboplatin resistant tumors unnecessary suffering and expense by avoiding an ineffective treatment, 3) allowing patients with carboplatin resistant tumors to pursue alternative treatment options without delay, and 4) providing a general model for identifying tumors resistant to other chemotherapeutic regimens. To achieve these goals, many genes must be evaluated. Carcinoma cell lines will be the most appropriate model for a comprehensive investigation. The specific aims are the following. Aim 1) Determine expression of eighty putative carboplatin chemoresistance genes in ten NSCLC lines, determine the IC30 concentration of carboplatin for each line, then determine which constitutive expression patterns correlate best statistically (by linear regression or by Duncan's Multiple Range test) with resistance. Any correlation identified will be validated in ten additional NSCLC lines. Aim 2) Combine the individual constitutive gene expression values into indices and assess them for association with the initial ten chemoresistant cell lines. Any correlation will be validated in the second ten lines. The competitive template RT-PCR gene expression quantification method proposed for this project is suggested by the applicant to be the best available because 1) it allows high throughput measurement of many samples. 2) the data produced are in standard units so that individual gene expression values may be combined mathematically in gene expression indices, and 3) because the same standard mixture of competitive templates will be used for all of the proposed experiments, data from all experiments may be compared for statistical analysis.