Inhibition of heat shock protein 90 function by 17-allylamino-17-demethoxy-geldanamycin in Hodgkin's lymphoma cells down-regulates Akt kinase, dephosphorylates extracellular signal-regulated kinase, and induces cell cycle arrest and cell death
Inhibition of heat shock protein 90 function by 17-allylamino-17-demethoxy-geldanamycin in Hodgkin's lymphoma cells down-regulates Akt kinase, dephosphorylates extracellular signal-regulated kinase, and induces cell cycle arrest and cell death
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DOI：
10.1158/1078-0432.ccr-05-1194
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发表时间：
2006-01-15
影响因子：
11.5
通讯作者：
Younes, A
中科院分区：
文献类型：
--
作者：
Georgakis, GV;Li, Y;Younes, A
目的：热休克蛋白90（HSP 90）是参与转录调节、信号转导和细胞周期控制的几种客户蛋白的伴侣。HSP 90在多种肿瘤类型中大量表达，最近已成为癌症治疗的靶点。本研究的目的是确定HSP 90在促进霍奇金淋巴瘤的生长和存活中的作用，并确定小分子17-烯丙基氨基-17-去甲氧基-格尔德霉素（17-AAG）在霍奇金淋巴瘤中抑制HSP 90功能的分子后果。HSP 90在霍奇金淋巴瘤细胞系中的表达通过蛋白质印迹法测定，在霍奇金淋巴瘤患者的原发淋巴结切片中的表达通过免疫组织化学测定。通过3-（4，5-二甲基-噻唑-2基）5-（3-羧基甲氧基苯基）-2-（4-磺基苯基）-2H-四唑鎓（MTS）测定法测定细胞活力。流式细胞仪检测细胞凋亡和细胞周期。结果：HSP 90在原代和培养的霍奇金淋巴瘤细胞中均呈高表达。17-AAG对HSP 90功能的抑制显示出对霍奇金淋巴瘤细胞系的时间和剂量依赖性生长抑制。17-AAG诱导细胞周期阻滞和凋亡，这与细胞周期蛋白依赖性激酶（CDK）4，CDK 6和polo样激酶1（PLK 1）的减少有关，并通过半胱天冬酶依赖性和非半胱天冬酶依赖性机制诱导凋亡。此外，17-AAG耗尽Akt的细胞内容物，降低细胞外信号调节激酶（ERK）磷酸化，并降低细胞FLICE样抑制蛋白水平（FLIP），从而增强多柔比星和激动性抗肿瘤坏死因子相关凋亡诱导配体（TRAIL）死亡受体抗体的细胞毒性作用。HSP 90功能的抑制诱导细胞死亡并增强化疗和抗肿瘤坏死因子相关凋亡诱导配体死亡受体抗体的活性，表明靶向HSP 90功能可能在霍奇金淋巴瘤中具有治疗价值。
Purpose: Heat shock protein 90 (HSP90) is a chaperone for several client proteins involved in transcriptional regulation, signal transduction, and cell cycle control. HSP90 is abundantly expressed by a variety of tumor types and has been recently targeted for cancer therapy. The objective of this study was to determine the role of HSP90 in promoting growth and survival of Hodgkin's lymphoma and to determine the molecular consequences of inhibiting HSP90 function by the small-molecule 17-allylamino-17-demethoxy-geldanamycin (17-AAG) in Hodgkin's lymphoma.Experimental Design: HSP90 expression in Hodgkin's lymphoma cell lines was determined by Western blot and in primary lymph node sections from patients with Hodgkin's lymphoma by immunohistochemistry. Cell viability was determined by the 3-(4,5-dimethyl-thiazol-2yl)5-(3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2H-tetrazolium (MTS) assay. Apoptosis and cell cycle fractions were determined by flow cytometry. Expression of intracellular proteins was determined by Western blot.Results: HSP90 is overexpressed in primary and cultured Hodgkin's lymphoma cells. Inhibition of HSP90 function by 17-AAG showed a time- and dose-dependent growth inhibition of Hodgkin's lymphoma cell lines. 17-AAG induced cell cycle arrest and apoptosis, which were associated with a decrease in cyclin-dependent kinase (CDK) 4, CDK 6, and polo-like kinase 1 (PLK1), and induced apoptosis by caspase-dependent and caspase-independent mechanisms. Furthermore, 17-AAG depleted cellular contents of Akt, decreased extracellular signal - regulated kinase (ERK) phosphorylation, and reduced cellular FLICE-like inhibitory protein levels (FLIP), and thus enhanced the cytotoxic effect of doxorubicin and agonistic anti - tumor necrosis factor - related apoptosis-inducing ligand (TRAIL) death receptor antibodies.Conclusion: Inhibition of HSP90 function induces cell death and enhances the activity of chemotherapy and anti - tumor necrosis factor - related apoptosis-inducing ligand death receptor antibodies, suggesting that targeting HSP90 function might be of therapeutic value in Hodgkin's lymphoma.