Activities of multiple cancer-related pathways are associated with BRAF mutation and predict the resistance to BRAF/MEK inhibitors in melanoma cells
Activities of multiple cancer-related pathways are associated with BRAF mutation and predict the resistance to BRAF/MEK inhibitors in melanoma cells
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DOI：
10.4161/cc.26971
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发表时间：
2014-01-15
期刊：
影响因子：
4.3
通讯作者：
Xing, Mingzhao
中科院分区：
文献类型：
--
作者：
Liu, Dingxie;Liu, Xuan;Xing, Mingzhao
耐药是使用BRAF/MEK抑制剂靶向治疗黑色素瘤的主要障碍。本研究旨在确定BRAF v600e相关的致癌途径，预测BRAF突变黑色素瘤对BRAF/MEK抑制剂的耐药性。我们采用计算机方法分析了黑色素瘤细胞中24种癌症相关通路的活性，并确定了那些激活与BRAF V600E相关的通路，并使用支持向量机（SVM）算法预测了BRAF突变的黑色素瘤细胞对BRAF/MEK抑制剂的耐药性。然后，我们通过实验证实了计算机上的发现。在63个黑色素瘤细胞系的微阵列基因表达数据集中，我们发现多种致癌途径的激活优先发生在braf突变的黑色素瘤细胞中。这一发现在另外5个独立的黑色素瘤数据集中得到了重复。对46个BRAF突变的黑色素瘤细胞系的进一步分析显示，与应答性黑色素瘤细胞相比，azd6244耐药细胞中TNF - α、EGFR、IFN - α、缺氧、IFN - γ、STAT3和MYC等7种通路的表达显著不同。在我们的实验中，基于这7通路激活模式构建的SVM分类器正确预测了10个braf突变的黑色素瘤细胞系对MEK抑制剂AZD6244的反应。我们通过实验发现，与子系DRO相比，黑色素瘤细胞A375中TNF α、EGFR、IFN α和IFN γ通路活性也上调，而DRO对AZD6244的敏感性远高于A375。总之，我们已经确定了在BRAF突变的黑色素瘤细胞中优先激活的特定致癌途径，以及预测BRAF突变的黑色素瘤对BRAF/MEK抑制剂耐药的途径模式，为黑色素瘤治疗提供了新的临床意义。
Drug resistance is a major obstacle in the targeted therapy of melanoma using BRAF/MEK inhibitors. This study was to identify BRAF V600E-associated oncogenic pathways that predict resistance of BRAF-mutated melanoma to BRAF/MEK inhibitors. We took in silico approaches to analyze the activities of 24 cancer-related pathways in melanoma cells and identify those whose activation was associated with BRAF V600E and used the support vector machine (SVM) algorithm to predict the resistance of BRAF-mutated melanoma cells to BRAF/MEK inhibitors. We then experimentally confirmed the in silico findings. In a microarray gene expression dataset of 63 melanoma cell lines, we found that activation of multiple oncogenic pathways preferentially occurred in BRAF-mutated melanoma cells. This finding was reproduced in 5 additional independent melanoma datasets. Further analysis of 46 melanoma cell lines that harbored BRAF mutation showed that 7 pathways, including TNF alpha, EGFR, IFN alpha, hypoxia, IFN gamma, STAT3, and MYC, were significantly differently expressed in AZD6244-resistant compared with responsive melanoma cells. A SVM classifier built on this 7-pathway activation pattern correctly predicted the response of 10 BRAF-mutated melanoma cell lines to the MEK inhibitor AZD6244 in our experiments. We experimentally showed that TNF alpha, EGFR, IFN alpha, and IFN gamma pathway activities were also upregulated in melanoma cell A375 compared with its sub-line DRO, while DRO was much more sensitive to AZD6244 than A375. In conclusion, we have identified specific oncogenic pathways preferentially activated in BRAF-mutated melanoma cells and a pathway pattern that predicts resistance of BRAF-mutated melanoma to BRAF/MEK inhibitors, providing novel clinical implications for melanoma therapy.