Curry Jonathan L, Torres-Cabala Carlos A, Tetzlaff Michael T, Bowman Christopher, Prieto Victor G
Department of Pathology, Section of Dermatopathology, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA.
Semin Cutan Med Surg. 2012 Dec;31(4):267-73. doi: 10.1016/j.sder.2012.07.007.
Metastatic melanoma (MM) is a deadly skin disease refractory to standard chemotherapy. Despite numerous clinical and pathological parameters derived to guide patient management, clinical outcomes in melanoma patients remain difficult to predict. There is a critical need to delineate the important biomarkers typical of this disease. These biomarkers will ideally illuminate those key biochemical pathways responsible for the aggressive behavior of melanoma and, in the process, unveil new opportunities for the design of rational therapeutic interventions in high-risk patients. The most common recurring mutation in cutaneous melanoma is the prooncogenic BRAF V600E mutation that drives melanoma cell proliferation. The development of RAF inhibitors targeted against BRAF V600E mutant melanoma cells has revolutionized the treatment of MM. Clinical trials with BRAF inhibitor vemurafenib have shown objective clinical response and improved survival in patients with MM; therefore, knowledge of the molecular signature of melanoma in patients will be important in directing management decisions. Several molecular platforms exist to analyze the mutation status of melanoma. These include Sanger sequencing, pyrosequencing, allele-specific reverse transcriptase polymerase chain reaction, mass spectrometry base sequencing (Sequenom), high-resolution melting curve analysis, and next-generation sequencing methods using microfluidics technology. The Food and Drug Administration has approved the cobas BRAF V600 Mutation Test developed by Roche to analyze BRAF mutation status in formalin-fixed paraffin-embedded tumor samples. The cobas Mutation Test has been designed specifically to detect BRAF V600E mutations, and the analytic performance of this assay has demonstrated >99% sensitivity in the detection of BRAF V600E mutation when compared with the Sanger sequencing method and confirmed with the next-generation sequencing 454-pyrosequencing technology. The lower limit of detection of the percentage of mutant alleles in a tissue sample for the cobas test is less than 4%-5%. Some cross-reactivity with other variants of mutant BRAF was seen with the cobas V600 platform; however, this clinical test offers highly sensitive reproducible BRAF V600E mutation analysis in formalin-fixed paraffin-embedded tumor samples.
转移性黑色素瘤（MM）是一种对标准化疗难治的致命性皮肤疾病。尽管有许多用于指导患者管理的临床和病理参数，但黑色素瘤患者的临床结局仍然难以预测。迫切需要确定这种疾病典型的重要生物标志物。理想情况下，这些生物标志物将阐明那些导致黑色素瘤侵袭性行为的关键生化途径，并在此过程中揭示针对高危患者设计合理治疗干预措施的新机会。皮肤黑色素瘤中最常见的复发性突变是致癌性BRAF V600E突变，它驱动黑色素瘤细胞增殖。针对BRAF V600E突变黑色素瘤细胞的RAF抑制剂的开发彻底改变了MM的治疗方法。使用BRAF抑制剂维莫非尼的临床试验已显示出MM患者有客观的临床反应并改善了生存率；因此，了解患者黑色素瘤的分子特征对于指导管理决策很重要。存在几种分子平台来分析黑色素瘤的突变状态。这些包括桑格测序、焦磷酸测序、等位基因特异性逆转录酶聚合酶链反应、质谱碱基测序（Sequenom）、高分辨率熔解曲线分析以及使用微流控技术 的下一代测序方法。美国食品药品监督管理局已批准罗氏公司开发的cobas BRAF V600突变检测法，用于分析福尔马林固定石蜡包埋肿瘤样本中的BRAF突变状态。cobas突变检测法专门设计用于检测BRAF V600E突变，与桑格测序法相比，该检测方法在检测BRAF V600E突变时的分析性能显示出>99%的灵敏度，并通过下一代测序454焦磷酸测序技术得到证实。cobas检测法在组织样本中检测突变等位基因百分比的检测下限小于4%-5%。在cobas V600平台上观察到与突变BRAF的其他变体有一些交叉反应；然而，这项临床试验在福尔马林固定石蜡包埋肿瘤样本中提供了高度敏感的可重复的BRAF V600E突变分析。
