GENE EXPRESSION INDICES ASSOCIATED WITH CHEMORESISTANCE

描述：（申请人的摘要）非小细胞肺癌（NSCLC）是 对目前的化疗方案反应较差， 回归率仅为30- 50%。抗药性的机制可能 涉及多种基因产物。在初步研究中，H1435非小细胞 肺癌（NSCLC）细胞系对卡铂的耐药性是 H460。20个耐药相关基因的检测 揭示谷胱甘肽转移酶（GST）P1、Bax α、GADD 45、ERCC 3和 谷胱甘肽过氧化物酶，mGST基因在100、20、10、6、5和100个氨基酸水平表达。 4-在H1435中的水平分别高一倍。的主要长期目标 拟开展的研究旨在提高对NSCLC机制的认识 化疗耐药性，并开发一种方法来预测哪些NSCLC肿瘤将 对卡铂有反应实现这些目标将有具体的价值1） 允许基于知识开发改进的化学治疗剂， 机制，2）保留卡铂耐药肿瘤患者 不必要的痛苦和费用，避免无效的治疗，3） 使卡铂耐药肿瘤患者能够寻求替代疗法， 治疗选择没有延误，和4）提供了一个通用的模式， 鉴定对其他化疗方案有抗性的肿瘤。实现 为了达到这些目标，必须对许多基因进行评估。癌细胞系将是 最适合进行全面调查的模式。具体目标是 下面的.目的1）确定80个推定的卡铂的表达 在10个NSCLC细胞系中的化疗耐药基因，确定IC 30浓度， 卡铂，然后确定哪种组成型表达 模式在统计学上相关性最好（通过线性回归或邓肯 多量程测试）。确定的任何相关性将 在另外10个NSCLC细胞系中进行了验证。目标2）联合收割机 组成型基因表达值转化为指数，并评估它们， 与最初的十种化疗耐药细胞系相关。任何相关性 将在后十行中验证。竞争性模板RT-PCR基因 表达量化的方法建议为这个项目是由 申请人是最好的，因为1）它允许高吞吐量 许多样品的测量。2)产生的数据是标准单位， 各个基因表达值可以在基因中数学组合 表达指数，和3），因为相同的标准混合的竞争 模板将用于所有拟议的实验，数据来自所有 可以比较实验以进行统计分析。

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.