Inhibition of HER-2 by three independent targeting strategies increases paclitaxel resistance of SKOV-3 ovarian carcinoma cells
Inhibition of HER-2 by three independent targeting strategies increases paclitaxel resistance of SKOV-3 ovarian carcinoma cells
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DOI：
10.1007/s00210-004-1016-4
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发表时间：
2005-02-01
影响因子：
3.6
通讯作者：
Aigner, A
中科院分区：
文献类型：
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作者：
Abuharbeid, S;Apel, J;Aigner, A
卵巢癌是最常见的妇科恶性肿瘤之一，目前的治疗选择有限，主要是因为晚期疾病患者往往会对化疗产生耐药性。在乳腺癌细胞中，多项研究表明，人表皮生长因子受体 2 (HER-2) 的过度表达会导致对某些（但并非所有）细胞毒性药物的耐药性增加。在卵巢癌中，关于 HER-2 表达与肿瘤细胞敏感性的相关性存在相互矛盾的数据。在本文中，我们通过在 SKOV-3 卵巢癌细胞中应用三种不同且独立的策略，探讨 HER-2 表达和信号水平与紫杉醇 (Taxol) 化疗耐药相关的作用。首先，我们发现，使用肿瘤治疗中成熟的 HER-2 抑制性抗体曲妥珠单抗（赫赛汀）治疗会导致细胞紫杉醇耐药性显着增加而不是降低。接下来，我们介绍两种新开发的 HER-2 酪氨酸激酶活性低分子量抑制剂：D-69491 和 D-70166。使用这两种药物，证实了 HER-2 抑制后细胞紫杉醇敏感性的降低。最后，为了进行更详细的分析，我们通过核酶靶向稳定下调 HER-2 表达。使用具有不同残留 HER-2 水平的克隆核酶转染的 SKOV-3 细胞，我们建立了紫杉醇细胞毒性的“HER-2 基因剂量效应”。我们证明这种作用是由于紫杉醇对细胞凋亡的差异诱导和细胞周期的差异抑制所致。最后，讨论了紫杉醇或 HER-2 介导的 MAP 激酶 p42/44、应激激活蛋白激酶/Jun 末端激酶 (SAPK/JNK) 和 p38 磷酸化的改变，以及对 caspase-3、caspase-7 和 bcl-2 激活的影响。我们得出结论，紫杉醇在 SKOV-3 细胞中的细胞毒性是“HER-2 剂量依赖性”，并将细胞增殖确定为这种效应的一个潜在细胞事件。
Current treatment options for ovarian cancer, which is one of the most widespread gynecological malignancies, are limited, mainly because patients with advanced-stage disease often develop resistance to chemotherapeutics. In breast cancer cells, several studies suggest that overexpression of the human epidermal growth factor receptor-2 (HER-2) leads to increased resistance against certain, but not all cytotoxic drugs. In ovarian carcinoma, conflicting data on the correlation of HER-2 expression and tumor cell sensitivity exist. In this paper, we explore the role of HER-2 expression and signaling levels pertaining to paclitaxel (Taxol) chemoresistance by applying three different and independent strategies in SKOV-3 ovarian carcinoma cells. Firstly, we show that treatment with the HER-2 inhibitory antibody trastuzumab (Herceptin), which is well established in tumor therapy, results in markedly increased, rather than decreased, cellular paclitaxel resistance. Next, we present two newly developed low molecular weight inhibitors of HER-2 tyrosine kinase activity, D-69491 and D-70166. With both drugs, the decrease in cellular paclitaxel sensitivity upon HER-2 inhibition is confirmed. Finally, for more detailed analysis we stably downregulate HER-2 expression by ribozyme-targeting. Using clonal ribozyme-transfected SKOV-3 cells with different residual HER-2 levels, we establish a 'HER-2 gene dose effect' of paclitaxel cytotoxicity. We show that this effect is due to differential induction of apoptosis and differential cell cycle inhibition by paclitaxel. Finally, paclitaxel- or HER-2-mediated alterations in the phosphorylation of MAP kinases p42/44, Stress-activated protein kinase/Jun-terminal kinase (SAPK/JNK), and p38, and effects on the activation of caspase-3, caspase-7, and bcl-2 are discussed. We conclude that paclitaxel cytotoxicity in SKOV-3 cells is 'HER-2 dose-dependent' and identify cell proliferation as one underlying cellular event of this effect.