The FA/BRCA pathway is involved in melphalan-induced DNA interstrand cross-link repair and accounts for melphalan resistance in multiple myeloma cells
The FA/BRCA pathway is involved in melphalan-induced DNA interstrand cross-link repair and accounts for melphalan resistance in multiple myeloma cells
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DOI：
10.1182/blood-2004-11-4286
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发表时间：
2005-07-15
期刊：
影响因子：
20.3
通讯作者：
Dalton, WS
中科院分区：
文献类型：
--
作者：
Chen, Q;Van der Sluis, PC;Dalton, WS
美法仑是一种 DNA 交联剂，是治疗多发性骨髓瘤 (MM) 中使用最广泛、最有效的药物之一。在本报告中，我们证明通过范可尼贫血 (FA)/BRCA 途径增强的链间交联 (ICL) 修复有助于马法兰耐药性骨髓瘤细胞系获得耐药性，而破坏该途径可逆转耐药性。使用碱性彗星测定（单细胞凝胶电泳），我们观察到与马法兰敏感细胞相比，马法兰耐药细胞减少了 ICL 形成并增强了 ICL 修复。细胞周期研究表明，增强的 ICL 修复可以使细胞免受美法仑诱导的细胞周期延迟的影响。使用 siRNA 敲低 8226/LR5 和 U266/LR6 耐药细胞中的 FANCF，证明 ICL 修复能力和药物敏感性之间存在直接关系。 FANCF在8226/S和U266/S药物敏感细胞中的过表达部分再现了耐药表型。这些数据表明，通过 Fanconi 贫血/BRCA 途径增强的 DNA 修复与获得性美法仑耐药有关。我们的研究结果为增强癌症治疗中 DNA 交联剂的反应提供了一个新的靶点。
Melphalan, a DNA cross-linker, is one of the most widely used and effective drugs in the treatment of multiple myeloma (MM). In this report, we demonstrate that enhanced interstrand cross-link (ICL) repair via the Fanconi anemia (FA)/BRCA pathway contributes to acquired drug resistance in melphalan-resistant myeloma cell lines, and disruption of this pathway reverses drug resistance. Using the alkaline comet assay (single-cell gel electrophoresis), we observed that melphalanresistant cells have reduced ICL formation and enhanced ICL repair compared with melphalan-sensitive cells. Cell-cycle studies demonstrated that enhanced ICL repair released cells from melphalan-induced cell-cycle delay. Using siRNA to knock down FANCF in 8226/LR5 and U266/ LR6 drug-resistant cells demonstrated a direct relationship between ICL repair capacity and drug sensitivity. Overexpression of FANCF in 8226/S and U266/S drug-sensitive cells partially reproduced the drug-resistant phenotype. These data show that enhanced DNA repair via the Fanconi anemia/BRCA pathway is involved in acquired melphalan resistance. Our findings provide for a new target to enhance response to DNA cross-linking agents in cancer treatment.