Protection of platinum-DNA adduct formation and reversal of cisplatin resistance by anti-MRP2 hammerhead ribozymes in human cancer cells
Protection of platinum-DNA adduct formation and reversal of cisplatin resistance by anti-MRP2 hammerhead ribozymes in human cancer cells
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DOI：
10.1002/ijc.20899
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发表时间：
2005-06-20
影响因子：
6.4
通讯作者：
Lage, H
中科院分区：
文献类型：
--
作者：
Materna, V;Liedert, B;Lage, H
含铂抗肿瘤药物的耐药性是其临床使用的主要限制。阐明ABC转运蛋白MRP2在铂类耐药中的作用。在人顺铂耐药细胞系：卵巢癌系 A2780RCIS、肾上腺皮质癌系 D43/86RCIS 和黑色素瘤系 MeWoCIS1 中分析了其表达。所有这些细胞均表现出 MRP2 的过度表达。为了逆转铂耐药性，将 2 个抗 MRP2 锤头核酶引入 A2780RCIS 细胞中。两种核酶都表现出基因沉默活性并逆转了耐药表型。此外，在DNA中测量了铂诱导的链内交联的形成。 DNA铂化水平与MRP2表达水平成反比，并伴随着caspase-3依赖性细胞凋亡的增加。铂-DNA 加合物的形成和消除动力学表明 DNA 修复能力没有改变；铂-DNA加合物形成的减少更多地反映了MRP2的保护活性。总之，MRP2 的功能性抑制可能是逆转铂类抗癌药物耐药性的一种有前景的策略。核酶技术对 MRP2 的特异性抑制反映了这一点，表明这种基因治疗方法可能适用于作为克服人类肿瘤铂耐药性的特定手段。 (c) 2005 年 Wiley-Liss, Inc.
Resistance to platinum-containing antineoplastic drugs is the major limitation in their clinical use. To elucidate the role of the ABC transporter MRP2 in platinum drug resistance,. its expression was analyzed in human cisplatin -resistant cell lines: the ovarian carcinoma line A2780RCIS, the adrenocortical carcinoma line D43/86RCIS and the melanoma line MeWoCIS1. All these cells showed overexpression of MRP2. For reversal of platinum resistance, 2 anti-MRP2 hammerhead ribozymes were introduced into A2780RCIS cells. Both ribozymes showed gene-silencing activities and reversed the drug-resistant phenotype. Moreover, formation of platinum -induced intrastrand cross-links was measured in DNA. The level of DNA platination corresponded inversely to the level of MRP2 expression and was accompanied by increased caspase-3-dependent apoptosis. Kinetics of formation and elimination of platinum-DNA adducts suggest that the DNA repair capacity was not altered; the decrease in platinum-DNA adduct formation was rather a reflection of the protecting activity of MRP2. In conclusion, functional inhibition of MRP2 might be a promising strategy in the reversal of resistance to platinum-based anticancer drugs. This was reflected by the specific inhibition of MRP2 by ribozyme technology, indicating that this gene therapeutic approach may be applicable as a specific means to overcome platinum resistance in human neoplasms. (c) 2005 Wiley-Liss, Inc.