Mecklenburg Ingo, Weckermann Dorothea, Zippelius Alfred, Schoberth Alexandra, Petersen Stephanie, Prang Nadja, Riethmüller Gert, Kufer Peter
Institute of Immunology, Ludwig-Maximilians-University, Munich, Germany.
J Immunol Methods. 2007 Jun 30;323(2):180-93. doi: 10.1016/j.jim.2007.04.006. Epub 2007 May 15.
Distant metastases of solid tumors are usually associated with fatal outcome. Disseminated cancer cells are considered early indicators of metastasis. Their sensitive detection and quantification would be a valuable tool for staging of disease and as guidance for therapeutic decisions.
We established a highly sensitive and quantitative multimarker real-time RT-PCR assay for amplification of cancer-related genes MAGE-A1, -A2, -A3/6, -A4, -A10 and -A12 using SYBR green I to detect one single tumor cell in 2 mL of blood or bone marrow. The feasibility of the assay was tested in a large cohort of 177 patients with locally confined prostate carcinoma.
Analysis revealed frequent MAGE expression in venous blood and bilateral bone marrow samples (25.5% of all cases) and yielded the first quantitative profile of MAGE expression with a broad range of transcript concentrations for individual markers in the minimal systemic tumor load of patients with localized cancer.
Rare transcripts of different MAGE-A genes can be quantified in clinical samples of cancer patients by a sensitive multimarker real-time RT-PCR. Because of frequent expression of MAGE genes in various types of cancer the multimarker MAGE real-time RT-PCR may be generally useful for detection, quantification and characterization of the individual disseminated tumor load in cancer patients.
实体瘤的远处转移通常与致命结局相关。播散癌细胞被认为是转移的早期指标。对其进行灵敏检测和定量将成为疾病分期及治疗决策指导的一项宝贵工具。
我们建立了一种高度灵敏的多标志物实时逆转录聚合酶链反应(RT-PCR)检测方法,用于扩增癌症相关基因MAGE-A1、-A2、-A3/6、-A4、-A10和-A12,使用SYBR绿I在2 mL血液或骨髓中检测单个肿瘤细胞。该检测方法的可行性在177例局限性前列腺癌患者的大型队列中进行了测试。
分析显示静脉血和双侧骨髓样本中MAGE频繁表达(占所有病例的25.5%),并得出了MAGE表达的首个定量图谱,在局限性癌症患者的最小全身肿瘤负荷中,各个标志物的转录本浓度范围广泛。
通过灵敏的多标志物实时RT-PCR可对癌症患者临床样本中不同MAGE-A基因的罕见转录本进行定量。由于MAGE基因在各种类型癌症中频繁表达,多标志物MAGE实时RT-PCR可能普遍有助于检测、定量和表征癌症患者个体的播散肿瘤负荷。