肿瘤相关基因RRM1和TUBB3 mRNA表达水平的检测在个体化用药中的初步应用

【作者】 王琪；

【导师】 陈超；

【作者基本信息】 西北大学 ， 生物化学与分子生物学， 2013， 硕士

【摘要】 随着药物基因组学的发展,个体化诊疗已成为临床疾病诊断治疗的关键。药物代谢在种族以及个体之间都存在着很大的差异,从而造成个体对药物的效应、毒性和耐药程度的不同。研究显示,这些差异大多是由个体遗传变异造成的。实现个体化治疗,可为特定人群设计最为有效的药物,提高疗效,缩短病程,减少毒副作用和成本。吉西他滨和紫杉醇是广谱的抗肿瘤药物,常被用于治疗肺癌、乳腺癌、胃癌、结直肠癌等多种肿瘤。大量临床研究表明,RRM1mRNA表达水平与吉西他滨治疗效果密切相关,RRM1是吉西他滨耐药的一个重要的预测指标,其mRNA过度表达会导致对吉西他滨化疗不敏感。TUBB3mRNA表达水平与紫杉醇治疗效果密切相关,其表达水平低的患者使用抗微管类药物进行治疗的效果更好。因此患者在使用吉西他滨或紫杉醇类药物化疗前,检测RRM1或TUBB3基因mRNA表达水平十分重要,可根据表达水平的高低预测治疗效果,辅助制定合理的治疗方案。本研究基于此对实验条件进行了摸索与优化,建立了重复性稳定性好、灵敏度高且操作简单的TaqMan探针荧光实时定量PCR检测RRM1/TUBB3mRNA表达水平的方法,用于指导临床吉西他滨及紫杉醇的个体化用药。通过已建立的检测方法检测了81例肿瘤样本,其中包括结直肠癌、乳腺癌、肺癌、胃癌在内的4种癌症患者的肿瘤样本中RRM1/TUBB3mRNA表达水平,分析了两种基因在不同肿瘤中表达的相关性,表达水平与年龄的相关性及在不同肿瘤中表达的差异性。结果表明RRM1、TUBB3基因mRNA表达情况与患者性别及年龄之间无相关性,且mRNA表达量均存在一定程度的组织差异性。更多还原

【Abstract】 With the development of pharmacogenomics, individualized treatment has become the key to the clinical diagnosis and treatment of disease. There exist great differences between race and individuals in metabolism, these differences may cause different effects in individual drugs, toxicity and resistance degree. The current study has shown that these differences are mainly caused by the differences of individual heritage. To achieve individual heritage, the most effective drugs could be designed to improve efficacy, shorten the course, reduce side effects and cost for specific populations.Gemcitabine and taxol is broad-spectrum anticancer drug, which have been widely used in the first-line treatment of non-small cell lung cancer, breast cancer, pancreatic cancer and bladder cancer. Numerous clinical studies have shown that the mRNA expression level of RRM1is closely related to the treatment effect of gemcitabine.The clinical data showed that RRM1was an important predictor of gemcitabine resistance, the mRNA over expression of which could cause non-sensitive effect to gemcitabine chemotherapy. The mRNA expression levels of TUBB3is closely related to the treatment effect of taxol, patients with low expression levels would obtain better therapeutic effect when anti-microtubule drugs have been used. Therefore the detection of mRNA expression level of RRM1or TUBB3before taking chemotherapy for patients who use gemcitabine or paclitaxel is very important, therapeutic effect could be predicted and development of rational treatment could be assisted according to the expression level.In order to guide clinical personalized medicine of gemcitabine and taxol through exploration and optimization of experimental conditions, detection of mRNA’s expression level with TaqMan probe real-time quantitative PCR assay method was proposed. This method is simple, convenient and has good stability and high sensitivity. The expression levels of RRM1and TUBB3of81tumor samples including colorectal cancer, breast cancer, lung cancer and gastric cancer were detected through established testing methods, the correlation of the expression level of two genes in different tumors and the correlation of expression of the two genes in different tumors and expression levels of different age and expression in different tumors were analyzed. The results showed that mRNA expression level had no correlation with gender and age of the patients and the mRNA expression levels had a certain degree of organizational differences.更多还原