外周血单个核细胞多药耐药相关基因含量监测在儿童恶性肿瘤中的应用  

Measuring multiple drug resistance related gene expression in the mononuclear cells of periphery blood in child malignant tumor

作　　者：谭俊杰[1] 张鹏辉[2] 邹琳[1] 

机构地区：[1]重庆医科大学附属儿童医院临床分子医学中心,重庆400014 [2]重庆医科大学附属儿童医院临床检验中心,儿童发育疾病研究省部共建教育部重点实验室,儿科学重庆市重点实验室,重庆市儿童发育重大疾病诊治与预防国际科技合作基地,重庆400014

出　　处：《重庆医科大学学报》2014年第1期1-4,共4页Journal of Chongqing Medical University

基　　金：国家自然科学基金资助项目(编号:90919013;30871103);重庆市自然科学基金资助项目(编号:2010BA5008);重庆市卫生局中医处基金资助项目(编号:2008-2-59)

摘　　要：目的:建立检测多药耐药(multi-drug resistance,MDR)相关基因荧光定量RT-PCR方法,并动态监测肿瘤患者外周血MDR相关基因(MDR1、MRP1和LRP)含量,探讨是否根据这些基因含量来指导肿瘤化疗。方法:利用Taq-Man探针方法,建立检测MDR相关基因的荧光定量RT-PCR方法。以健康对照组50例,肿瘤(含淋巴瘤、神经母细胞瘤、卵黄囊瘤等)组262例,化疗不同时期抽取患者外周血950份,提取RNA,检测单个核细胞内MDR相关基因(MDR1、MRP1和LRP)含量,分析这些基因的含量与肿瘤化疗方案的关系。结果:成功建立以Taq-Man探针为基础的MDR相关基因荧光定量RT-PCR方法。①与健康对照组相比,肿瘤组患者外周血单个核细胞(peripheral blood mononuclear,PBMC)MDR1基因起始含量较高,尤以神经母细胞瘤和卵黄囊瘤为甚,这2类肿瘤可能存在一定程度的原发耐药性。②对同一肿瘤患者,化疗后PBMC MDR1含量随化疗疗程增加而逐渐增高,以神经母细胞瘤和卵黄囊瘤最为敏感。结论:MDR1、MRP1和LRP含量与肿瘤的化疗方案密切相关,PBMC和肿瘤组织中MDR1的表达与化疗疗效呈显著负相关。检测外周血中MDR1基因的含量能够动态监测肿瘤患者的MDR,利用荧光定量RT-PCR方法动态监测肿瘤患者化疗前后外周血MDR相关基因(MDR1、MRP1和LRP)的含量,对化疗疗效监测及预后判定具有一定的参考价值,可以用来指导相关基因底物的肿瘤药物化疗。Objective：To establish the fluorescence quantitative RT-PCR method to measure multiple drug resistance （MDR） related genes in patient＇s periphery blood and to explore the feasibility of tumor chemotherapy instruction by dynamically detecting these genes in blood of patients with tumor. Methods ：Taq-Man probe was applied to establish the fluorescence quantitative RT-PCR method to measure MDR related genes. Periphery blood was collected from 262 patients with tumor（including lymphoma, neuroblsatoma, yolk sac tumor, etc.） and 50 healthy persons were taken as normal controls. Mononuclear cells were isolated from these samples,then RNAs were purified and expression of MDR related genes（MDR1, MRP1 and LRP） was detected. Relationship between gene expression and tumor chemotherapy was analyzed. Results：Fluorescence quantitative RT-PCR method measuring MDR related genes was established successfully based on Taq-Man probe. Compared with that in normal control group,expression of MDR related genes（MDR1 ,MRP1 and LRP） was increased in mononuclear cells of periphery blood of patients with tumor. For one tumor patient,expression of MDR relat- ed genes（MDR1, MRP1 and LRP） was higher in mononuclear cells of post-chemotherapy than in those of pre-chemotherapy. Expression of MDR1, MRP1 and LRP was closely related with tumor chemotherapy scheme of patients. Conclusions：Expression of MDR related genes（MDR1, MRP1 and LRP） in periphery blood of tumor patients is dynamically measured by fluorescence quantitative RT-PCR method,which could instruct the gene related substrate chem- otherapeutic drug.

关 键 词：多药耐药 外周血 荧光定量RT-PCR 儿童肿瘤 

分 类 号：R737.7[医药卫生—肿瘤]