αvβ3 integrin-mediated drug resistance in human laryngeal carcinoma cells is caused by glutathione-dependent elimination of drug-induced reactive oxidative species
αvβ3 integrin-mediated drug resistance in human laryngeal carcinoma cells is caused by glutathione-dependent elimination of drug-induced reactive oxidative species
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DOI：
10.1124/mol.107.043836
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发表时间：
2008-07-01
影响因子：
3.6
通讯作者：
Ambriovic-Ristov, Andreja
中科院分区：
文献类型：
--
作者：
Brozovic, Anamaria;Majhen, Dragomira;Ambriovic-Ristov, Andreja
作为确定整合素在耐药性中作用的模型，我们使用了整合素阴性的人喉癌细胞系(HEp2)和三个表达逐渐增加的整合素α(V)β(3)整合素的细胞克隆。α(V)β(3)整合素的表达保护细胞免受顺铂、丝裂霉素C和阿霉素的影响。在HEp2-α(V)β(3)整合素表达细胞中，Bcl2蛋白的组成性表达和谷胱甘肽(GSH)水平均高于HEp2细胞。用GSH合成抑制剂丁硫氨酸亚磺胺(BSO)处理HEp2-α(V)β(3)整合素表达细胞，可降低GSH水平，部分逆转对上述药物的耐药性，但不影响Bcl2的表达。HEp2细胞对某些抗癌药物的敏感性不随细胞中Bcl2的过度表达而改变，在表达整合素的HEp2α(V)β(3)细胞中也不随Bcl2的沉默而改变，提示Bcl2不参与耐药机制。HEp2和HEp2-α(V)β(3)整合素表达细胞DNA平台化差异无统计学意义，提示其耐药机制不依赖于GSH对顺铂的解毒作用。在HEp2-α(V)β(3)整合素表达细胞中，在顺铂或阿霉素处理期间HEp2细胞中反应性氧化物种(ROS)的形成显著增加。由于这种增加的ROS消除可以被GSH耗竭逆转，我们得出结论，多药耐药是GSH依赖的表达α(V)β(3)的细胞消除药物诱导的ROS能力增强的结果。
As a model for determination of the role of integrins in drug resistance, we used alpha(v)beta(3) integrin-negative human laryngeal carcinoma cell line (HEp2) and three HEp2-derived cell clones with a gradual increase of alpha(v)beta(3) integrin expression. The alpha(v)beta(3) integrin expression protects cells from cisplatin, mitomycin C, and doxorubicin. In HEp2-alpha(v)beta(3) integrin-expressing cells, the constitutive expression of Bcl-2 protein and the level of glutathione (GSH) were increased compared with HEp2 cells. Pretreatment of HEp2-alpha(v)beta(3) integrin-expressing cells with an inhibitor of GSH synthesis, buthionine sulfoximine (BSO), decreased the level of GSH and partially reverted drug resistance to all above-mentioned drugs, but it did not influence the expression of Bcl-2. Sensitivity to selected anticancer drugs did not change with overexpression of Bcl-2 in HEp2 cells, nor with silencing of Bcl-2 in HEp2-alpha(v)beta(3) integrin-expressing cells, indicating that Bcl-2 is not involved in resistance mechanism. There was no difference in DNA platination between HEp2 and HEp2-alpha(v)beta(3) integrin-expressing cells, indicating that the mechanism of drug resistance is independent of cisplatin detoxification by GSH. A strong increase of reactive oxidative species (ROS) formation during cisplatin or doxorubicin treatment in HEp2 cells was reduced in HEp2-alpha(v)beta(3) integrin-expressing cells. Since this increased elimination of ROS could be reverted by GSH depletion, we concluded that multidrug resistance is the consequence of GSH-dependent increased ability of alpha(v)beta(3)-expressing cells to eliminate drug-induced ROS.