HSP90 inhibitor 17-AAG plays an important role in JAK3/STAT5 signaling pathways in HTLV-1 infection cell line HUT-102

Qingqing Yang; Huo Tan; Zhiping Fu; Qiang Ma; Jinlong Song

doi:10.3760/cma.j.issn.0253-2727.2017.08.012

. 2017 Aug;38(8):710–715. [Article in Chinese] doi: 10.3760/cma.j.issn.0253-2727.2017.08.012

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HSP90 inhibitor 17-AAG plays an important role in JAK3/STAT5 signaling pathways in HTLV-1 infection cell line HUT-102

Qingqing Yang ¹, Huo Tan ^1,^✉, Zhiping Fu ¹, Qiang Ma ¹, Jinlong Song ¹

Editor: 徐茂强¹

PMCID: PMC7348253 PMID: 28954352

Abstract

Objective

To analyze whether heat-shock protein 90 (HSP90) be involved in a permanently abnormal activated JAK/STAT signaling in ATL cells in vitro.

Methods

The effect of 17-AAG on proliferation of ATL cell lines HUT-102 was assessed using CCK8 at different time points. Cell apoptosis was measured by flow cytometry. The specific proteins HSP90, STAT5, p-STAT5 and JAK3 were detected by Western blotting.

Results

Overexpression of HSP90 in HUT-102 cell lines was disclosed (P<0.05), and constitutive activation of JAK3/STAT5 signaling was observed in HTLV-1-infected T-cell lines but not in normal PBMCs; Treatment of ATL cell lines with 17-AAG led to reduced cell proliferation, but there was no significant change in terms of cell proliferation when the concentration of 17-AAG between 2 000–8 000 nmol/L (P>0.05). 17-AAG induced cell apoptosis in different time-points and concentrations. 17-AAG don't affect the expression of JAK3 gene.

Conclusion

This study indicated that JAK3 as HSP90 client protein was aberrantly activated in HTLV-1-infected T-cell lines, leading to constitutive activation of p-STAT5 in JAK/STAT signal pathway, which demonstrated that HSP90-inhibitors 17-AAG inhibited the growth of HTLV-1-infected T-cell lines by reducing cell proliferation and inducing cell apoptosis.

Keywords: Leukemia, T cell; HSP90 heat-shock proteins; Inhibitor

成人T细胞白血病（adult T-cell leukemia，ATL）是一种与人T细胞白血病病毒Ⅰ（HTLV-Ⅰ）感染直接相关、发生于成人的特殊类型淋巴系统恶性克隆增殖性疾病。急性期ATL侵袭性极强，其中位生存期为6~13个月，目前尚无标准治疗方案[1]。ATL细胞内存在多条信号通路的异常激活[2]–[3]，其中包括对细胞增殖和凋亡起到非常重要作用的JAK/STAT通路[4]。有研究指出，导致ATL细胞中JAK/STAT通路的异常持续性激活的因素并非来自胞外配体[5]。热休克蛋白（HSP）广泛存在于各种生物体细胞胞质内，参与翻译后初始多肽类的折叠与包装使之形成功能性蛋白，与细胞中一些功能蛋白如蛋白酶、类固醇激素受体等形成复合物并维持细胞内蛋白构象稳定与功能正常，帮助细胞在应激环境刺激下正常生长，参与细胞免疫和信号转导等一系列生命过程[6]–[8]。研究表明，ATL细胞内多条信号转导通路相关蛋白均为HSP90的客体蛋白，并通过与HSP90结合被异常激活[9]–[12]。本研究中，我们使用特异性HSP90抑制剂17-烯丙胺-17-脱甲氧格尔德霉素（17-AAG）探究ATL细胞内JAK/STAT信号通路的异常持续激活与HSP90蛋白的相关性。

材料与方法

1．主要试剂：17-AAG购自瑞士Alexis公司，以DMSO配成1 µmol/ml母液；RPMI 1640培养基和胎牛血清购自美国Gibco公司；兔抗酪氨酸激酶JAK3、信号传导及转录激活因子5（STAT5）、磷酸化STAT5（p-STAT5）、HSP90一抗购自美国CST公司，羊抗兔二抗购自美国KPL公司；CCK-8试剂盒购自日本同仁化学研究所；Annexin Ⅴ-FITC细胞凋亡检测试剂盒、TRIzol试剂、逆转录试剂盒均购自美国Invitrogen公司；引物由Invitrogen公司合成，分装后−20 °C保存；RIPA蛋白提取液购自上海申能博彩公司；碘化丙锭（PI）、蛋白酶抑制剂cocktail购自美国Sigma公司。

2．细胞培养：选用HTLV-1感染ATL细胞株HUT-102购自中国典型培养物保藏中心，普通人T淋巴细胞白血病细胞株Jurkat购自美国典藏物保存中心（ATCC）。正常人外周血单个核细胞（PBMC）提取自健康志愿者外周血标本。细胞重悬于含10%胎牛血清的完全培养基内，置于37 °C、5%CO₂培养箱培养。每2~3 d更换培养基。

3．CCK-8法检测不同浓度17-AAG对HUT-102细胞的增殖抑制作用：以不同浓度17-AAG（0、500、1 000、2 000、4 000、5 000、8 000 nmol/L）作用于对数生长期HUT-102细胞，分别于12、24、48、72 h后加入CCK-8试剂10 µl，在培养箱内继续孵育4 h，用酶标仪检测450 nm处各孔的吸光度（A）值。

4．流式细胞术检测不同浓度17-AAG对HUT-102细胞凋亡的影响：取对数生长期HUT-102细胞，分别将0、250、500、1 000、2 000 nmol/L 17-AAG加入细胞悬液内，并设置空白对照组，培养48 h后取出，收集各孔内细胞，使用AnnexinⅤ-PI试剂盒进行流式细胞术检测。

5．Western blot法检测细胞内蛋白表达：收集各组培养细胞，用含1 mmol/L苯甲基磺酰氟的RIPA裂解液裂解细胞，提取总蛋白，BCA试剂盒测总蛋白浓度，−80 °C保存。等量蛋白通过SDS-PAGE 2 h后电转移至聚偏氟乙烯（PVDF）膜，50 g/L脱脂奶粉溶液封闭1 h后加入一抗单抗、二抗多抗，用增强化学发光（ECL）试剂盒显色，扫描。

6．RT-PCR法检测细胞JAK3 mRNA表达：0、125、250、500、1000 nmol/L 17-AAG刺激HUT-102细胞24 h后，TRIzol法提取细胞总RNA，测定样品浓度及纯度。反应体系：5×reaction buffer 4 µl，RNase inhibitor 1 µl，10 mmol/L dNTPmix 2 µl，逆转录酶1 µl。反应条件：42 °C 60 min，70 °C灭活5 min。JAK3引物：上游5′-CAGCTGGGCAAGGGCAA CTTT-3′，下游5′-ACACCA CGATACTTGACAATG-3′；GAPDH引物：上游5′-GCCAGCCGAGCCACAT-3′，下游5′-CTTTACCAGAGTTAAAAGCAGCCC-3′，反应体系：Taq mix 12.5 µl，cDNA 2 µl，上游引物1 µl，下游引物1 µl，双蒸水8.5 µl。反应条件：98 °C变性10 s，63 °C退火30 s，72 °C延伸30 s，40次循环。扩增产物行20 g/L琼脂糖凝胶电泳（120 V，30 min）。于凝胶成像分析仪器上成像分析。

7．JAK3抑制剂CP690550联合17-AAG对HUT-102细胞株JAK3蛋白表达的影响：分别设立无刺激组、17-AAG+CP690550组、17-AAG组、CP690550组，作用于对数生长期HUT-102细胞24 h后收集细胞并提取总蛋白，Western blot检测JAK3蛋白表达情况。

8．统计学处理：实验数据利用SPSS13.0统计软件进行单因素方差分析，所有实验均重复3次以上，计量资料采用x±s表示，两组均数比较先进行方差齐性检验，方差齐者用t检验，方差不齐者用t ′检验，P<0.05认为差异有统计学意义。

结果

1．Western blot检测HSP90、JAK3、STAT5蛋白在正常人PBMC及HUT-102、Jurkat细胞中的表达：HUT-102以及Jurkat细胞株内HSP90蛋白以及总STAT5蛋白表达量显著高于正常人PBMC（P<0.05）。JAK3与p-STAT5表达情况有所不同，HUT-102细胞高表达JAK3与p-STAT5蛋白，而正常人PBMC以及Jurkat细胞几乎不表达（图1）。后续实验均选用高表达JAK3/p-STAT5的HUT-102细胞株。

2．CCK8方法检测17-AAG对HUT-102细胞的增殖抑制作用：17-AAG对HUT-102细胞的增殖抑制作用在一定范围内呈剂量与时间相关性（表1）。相同作用时间下，HUT-102细胞增殖活性随17-AAG浓度递增而逐渐下降,当17-AAG浓度达到2 000 nmol/L时细胞增殖活性差异无统计学意义（P>0.05）。当17-AAG浓度相同时，逐渐延长作用时间，细胞增殖活性亦逐渐下降，但48 h细胞增殖活性与72 h相比，差异无统计学意义（P>0.05）。详见表1。

表1. 17-烯丙胺-17-脱甲氧格尔德霉素（17-AAG）对HUT-102细胞增殖活性的影响（吸光度值，x±s）.

组别	12 h	24 h	48 h	72 h
空白对照	0.121±0.003	0.118±0.002	0.134±0.003	0.124±0.002
DMSO	1.068±0.002	1.454±0.003	1.749±0.002	1.513±0.001
0 nmol/L 17-AAG	1.031±0.003	1.472±0.002	1.582±0.005	1.337±0.004
500 nmol/L 17-AAG	0.856±0.002^a	0.658±0.001^a	0.481±0.004^a	0.324±0.005^a
1 000 nmol/L17-AAG	0.764±0.002^a	0.542±0.004^a	0.327±0.003^a	0.271±0.001^a
2 000 nmol/L 17-AAG	0.687±0.003^a	0.519±0.003^a	0.233±0.002^a	0.223±0.004^a
4 000 nmol/L 17-AAG	0.711±0.003^a	0.514±0.005^a	0.221±0.002^a	0.212±0.001^a
6 000 nmol/L 17-AAG	0.693±0.003^a	0.533±0.004^a	0.238±0.001^a	0.243±0.001^a
8 000 nmol/L 17-AAG	0.703±0.003^a	0.541±0.003^a	0.237±0.001^a	0.244±0.003^a

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注：与DMSO组比较，^aP<0.05

3．17-AAG对HUT-102细胞凋亡的影响：为了验证HSP90蛋白异常高表达对细胞凋亡的影响，设定0、250、500、1 000、2 000 nmol/L 5个17-AAG浓度组，分别作用于HUT-102细胞株48 h后检测细胞凋亡（同时设置空白对照）。未刺激组（0 nmol/L17-AAG）细胞凋亡率为（9.23±0.67）%，刺激组凋亡率随着17-AAG浓度增加而显著增加，浓度2 000 nmol/L时凋亡率达到（87.82±2.46）%，高于未刺激组（P<0.05）。

4．Western blot检测不同浓度17-AAG对HUT-102细胞JAK3/STAT5蛋白表达的抑制作用：125、250、500、1 000、2 000 nmol/L 17-AAG分别作用24、48 h后，HUT-102细胞JAK3与p-STAT5蛋白表达均低于未刺激组（0 nmol/L 17-AAG）（图2）。500、1 000、2 000 nmol/L浓度组作用24 h的蛋白表达水平低于未刺激组。在作用48 h时，p-STAT5蛋白表达低于未刺激组（P<0.01）。相同条件下，同一作用时间各浓度组之间总STAT5蛋白表达差异无统计学意义（P>0.05）。

A：17-AAG作用24 h；B：17-AAG作用48 h；1~6分别为0、125、250、500、1 000、2 000 nmol/L 17-AAG组

5．Western blot检测17-AAG作用不同时间对HUT-102细胞JAK3/STAT5蛋白表达的抑制作用：1 000 nmol/L 17-AAG作用0、3、6、9、12、24 h后，HUT-102细胞蛋白表达均下调。随着作用时间延长，JAK3蛋白表达水平逐渐降低，各组间差异有统计学意义（P<0.05）。在17-AAG作用≥3 h后p-STAT5蛋白均无明显表达，但总STAT5蛋白表达差异无统计学意义（P>0.05）。详见图3。

6．17-AAG对HUT-102细胞JAK3 mRNA表达的影响：0、250、500、1 000 nmol/L 17-AAG刺激HUT-102细胞24 h后，各组HUT-102细胞JAK3 mRNA表达与未加抑制剂组比较差异无统计学意义（图4），说明17-AAG未影响JAK3基因表达。

1、2、3、4分别为0、250、500、1 000 nmol/L 17-AAG组

7．JAK3抑制剂CP690550联合17-AAG对HUT-102细胞株JAK3蛋白表达的影响：如图5所示，17-AAG对JAK3蛋白抑制作用强于CP690550，17-AAG+CP690550共同刺激组细胞JAK3蛋白表达受抑明显强于单独刺激组，说明CP690550与17-AAG对HUT-102细胞JAK3蛋白表达的抑制可能存在协同作用。

讨论

实验证明，HSP90对肿瘤的发生发展起到十分重要作用，HSP90与客体蛋白形成的超级分子伴侣结构被认为是HSP90发挥促肿瘤形成的关键。特异性抑制HSP90在肿瘤细胞内的表达能显著诱导细胞凋亡、抑制细胞增殖，并且同时亦显著抑制众多细胞信号转导通路的异常激活[9],[12]–[18]。

Bcr-Abl、FLT-3、c-KIT、c-Raf、AKT等已被证实为HSP90的客体蛋白[17],[19]–[21]。有实验证明，通过抑制HSP90对客体蛋白的负调控作用强于直接抑制客体蛋白的表达[22]。在实体瘤以及髓性白血病体外实验中证明，JAK/STAT通路的异常持续性激活也与HSP90相关[23]。17-AAG与HSP90 N末端结构域ATP/ADP结合位点特异性竞争结合，抑制HSP90活性，并使超级分子伴侣复合物结构解体，使被HSP90异常持续活化的客体蛋白表达恢复正常[21]。

本实验以HTLV-1感染ATL细胞株HUT-102为研究对象，选择ATL细胞表达水平最高的JAK3、STAT5蛋白作为JAK/STAT信号通路的代表信号转导分子进行一系列体外实验研究，试图进一步了解HSP90与JAK/STAT通路之间的作用关系。

本研究中，我们利用Western blot技术对HTLV（+）T细胞株HUT-102、HTLV（−）细胞株Jurkat以及正常人PBMC进行细胞蛋白表达分析，结果显示正常人HSP90蛋白表达量显著低于两种T淋巴肿瘤细胞株，JAK3/STAT5蛋白几乎不表达。而与Jurkat细胞相比，HUT-102细胞内HSP90、JAK3、STAT5以及p-STAT5蛋白表达显著上调。提示恶性淋巴瘤细胞株内HSP90蛋白水平异常增加，HTLV（+）ATL细胞株内JAK3/STAT5通路异常持续性活化。HAP90特异性抑制剂17-AAG能显著抑制细胞增殖，诱导细胞凋亡，并且在一定范围内呈时间、剂量相关性。在上述实验中我们证明了HUT-102细胞内与增殖及凋亡密切相关的JAK3/STAT5通路异常活化，那么17-AAG对细胞增殖及凋亡的影响是否与JAK3/STAT5通路有关？首先我们我们以不同浓度17-AAG作用于HUT-102细胞不同时间并检测胞内JAK3/STAT5蛋白水平，结果发现17-AAG可以显著降低细胞内JAK3以及p-STAT5蛋白水平的表达，并且在一定浓度范围内（≤2 000 nmol/L），JAK3/p-STAT5蛋白水平随17-AAG浓度增加而逐渐下降。同样实验也证明了17-AAG对JAK3/p-STAT5蛋白水平的负调控呈时间相关性。在一定范围内随着时间的延长，17-AAG的作用强度逐渐增加。因此我们推测JAK3/STAT5通路的异常持续活化与HSP90的过高表达有一定的关系。在本研究中，我们还发现，无论17-AAG的浓度以及作用时间如何变化，细胞总STAT5蛋白表达水平变化不大，提示17-AAG只抑制p-STAT5蛋白表达。而由JAK/STAT通路的信号转导机制可知，由于JAK3激酶的激活，催化其下游STAT5蛋白磷酸化修饰，形成p-STAT5进一步在核内完成信号转导过程。由此可见，17-AAG对p-STAT5的抑制作用是通过抑制其上游的JAK3激酶来实现的，说明被17-AAG特异性抑制的HSP90蛋白与JAK3激酶之间存在某种相互作用。HSP90是通过影响其一种或多种客体蛋白进而影响JAK3蛋白的表达还是JAK3本身作为HSP90客体蛋白被异常激活，我们目前正在继续进行相关研究。

此前大多数的研究均表明，17-AAG通过直接作用于HSP90蛋白的ATP结合位点抑制HSP90的活性，并促使HSP90蛋白自身降解，从而下调肿瘤细胞HSP90蛋白的异常表达[24]–[28]。根据17-AAG的作用机制，我们利用不同浓度17-AAG刺激细胞，并检测细胞内JAK3 mRNA表达。结果显示，不同浓度刺激下的细胞JAK3 mRNA表达水平无显著差异，说明17-AAG JAK3的转录水平没有影响，也进一步证明HSP90与JAK3之间的关系可能存在于蛋白水平。

随后，我们进行JAK3特异性抑制剂CP690550与17-AAG共抑制实验，在17-AAG与CP690550共刺激作用下，JAK3蛋白表达量较两种抑制剂分别单独刺激下的JAK3表达量有所下降，提示这两种抑制剂在HUT-102细胞中可能存在协同作用。单独刺激下，17-AAG作用明显强于CP690550，可能的原因是在HUT-102细胞内存在多种HSP90客体蛋白的异常激活，抑制HSP90后除JAK3/STAT5通路出现负调控外，多种与HSP90客体蛋白相关的信号通路出现负调控所致[12]。这也进一步说明HSP90对ATL肿瘤发生发展的重要性。

本研究证明了在HTLV感染的ATL细胞株HUT-102细胞中，JAK3激酶作为HSP90的客体蛋白被异常活化，导致其下游信号蛋白p-STAT5异常持续性的激活。同时证明HSP90抑制剂17-AAG能显著下调JAK3、p-STAT5蛋白表达，抑制HUT-102细胞的增殖，诱导细胞的凋亡，并进一步证明JAK3特异性抑制剂与17-AAG之间可能存在协同作用，为今后开发多种靶向药物联合治疗提供了实验依据。HSP90与JAK3的关系尚需进行深入研究。

Funding Statement

基金项目：广州医学院第一附属医院科研基金青年项目（201224-gyfyy）

Fund program: The First Affiliated Hospital of Guangzhou Medical University Youth Program of Research Foundation（201224-gyfyy）

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6.Rappa F, Farina F, Zummo G, et al. HSP-molecular chaperones in cancer biogenesis and tumor therapy: an overview[J] Anticancer Res. 2012;32(12):5139–5150. [PubMed] [Google Scholar]
7.Joly AL, Wettstein G, Mignot G, et al. Dual role of heat shock proteins as regulators of apoptosis and innate immunity[J] J Innate Immun. 2010;2(3):238–347. doi: 10.1159/000296508. [DOI] [PubMed] [Google Scholar]
8.Soga S, Akinaga S, Shiotsu Y. Hsp90 inhibitors as anti-cancer agents, from basic discoveries to clinical development[J] Curr Pharm Des. 2013;19(3):366–376. doi: 10.2174/1381612811306030366. [DOI] [PubMed] [Google Scholar]
9.Broemer M, Krappmann D, Scheidereit C. Requirement of Hsp90 activity for IkappaB kinase (IKK) biosynthesis and for constitutive and inducible IKK and NF-kappaB activation[J] Oncogene. 2004;23(31):5378–5386. doi: 10.1038/sj.onc.1207705. [DOI] [PubMed] [Google Scholar]
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11.Fujita N, Sato S, Ishida A, et al. Involvement of Hsp90 in signaling and stability of 3-phosphoinositide-dependent kinase-1[J] J Biol Chem. 2002;277(12):10346–10353. doi: 10.1074/jbc.M106736200. [DOI] [PubMed] [Google Scholar]
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17.Yao Q, Nishiuchi R, Li Q, et al. FLT3 expressing leukemias are selectively sensitive to inhibitors of the molecular chaperone heat shock protein 90 through destabilization of signal transduction-associated kinases[J] Clin Cancer Res. 2003;9(12):4483–4493. [PubMed] [Google Scholar]
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22.Rao R, Nalluri S, Fiskus W, et al. Heat shock protein 90 inhibition depletes TrkA levels and signaling in human acute leukemia cells[J] Mol Cancer Ther. 2010;9(8):2232–2242. doi: 10.1158/1535-7163.MCT-10-0336. [DOI] [PMC free article] [PubMed] [Google Scholar]
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[b1] 1.Yamada Y, Tomonaga M, Fukuda H, et al. A new G-CSF-supported combination chemotherapy, LSG15, for adult T-cell leukaemia-lymphoma: Japan Clinical Oncology Group Study 9303[J] Br J Haematol. 2001;113(2):375–382. doi: 10.1046/j.1365-2141.2001.02737.x. [DOI] [PubMed] [Google Scholar]

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[b4] 4.Liongue C, O'Sullivan LA, Trengove MC, et al. Evolution of JAK-STAT pathway components: mechanisms and role in immune system development[J] PLoS One. 2012;7(3):e32777. doi: 10.1371/journal.pone.0032777. [DOI] [PMC free article] [PubMed] [Google Scholar]

[b5] 5.Migone TS, Lin JX, Cereseto A, et al. Constitutively activated Jak-STAT pathway in T cells transformed with HTLV-I[J] Science. 1995;269(5220):79–81. doi: 10.1126/science.7604283. [DOI] [PubMed] [Google Scholar]

[b6] 6.Rappa F, Farina F, Zummo G, et al. HSP-molecular chaperones in cancer biogenesis and tumor therapy: an overview[J] Anticancer Res. 2012;32(12):5139–5150. [PubMed] [Google Scholar]

[b7] 7.Joly AL, Wettstein G, Mignot G, et al. Dual role of heat shock proteins as regulators of apoptosis and innate immunity[J] J Innate Immun. 2010;2(3):238–347. doi: 10.1159/000296508. [DOI] [PubMed] [Google Scholar]

[b8] 8.Soga S, Akinaga S, Shiotsu Y. Hsp90 inhibitors as anti-cancer agents, from basic discoveries to clinical development[J] Curr Pharm Des. 2013;19(3):366–376. doi: 10.2174/1381612811306030366. [DOI] [PubMed] [Google Scholar]

[b9] 9.Broemer M, Krappmann D, Scheidereit C. Requirement of Hsp90 activity for IkappaB kinase (IKK) biosynthesis and for constitutive and inducible IKK and NF-kappaB activation[J] Oncogene. 2004;23(31):5378–5386. doi: 10.1038/sj.onc.1207705. [DOI] [PubMed] [Google Scholar]

[b10] 10.Villarino AV, Kanno Y, Ferdinand JR, et al. Mechanisms of Jak/STAT signaling in immunity and disease[J] J Immunol. 2015;194(1):21–27. doi: 10.4049/jimmunol.1401867. [DOI] [PMC free article] [PubMed] [Google Scholar]

[b11] 11.Fujita N, Sato S, Ishida A, et al. Involvement of Hsp90 in signaling and stability of 3-phosphoinositide-dependent kinase-1[J] J Biol Chem. 2002;277(12):10346–10353. doi: 10.1074/jbc.M106736200. [DOI] [PubMed] [Google Scholar]

[b12] 12.Kamal A, Thao L, Sensintaffar J, et al. A high-affinity conformation of Hsp90 confers tumour selectivity on Hsp90 inhibitors[J] Nature. 2003;425(6956):407–410. doi: 10.1038/nature01913. [DOI] [PubMed] [Google Scholar]

[b13] 13.Fujiwara H, Yamakuni T, Ueno M, et al. IC101 induces apoptosis by Akt dephosphorylation via an inhibition of heat shock protein 90-ATP binding activity accompanied by preventing the interaction with Akt in L1210 cells[J] J Pharmacol Exp Ther. 2004;310(3):1288–1295. doi: 10.1124/jpet.104.065979. [DOI] [PubMed] [Google Scholar]

[b14] 14.Pittet JF, Lee H, Pespeni M, et al. Stress-induced inhibition of the NF-kappaB signaling pathway results from the insolubilization of the IkappaB kinase complex following its dissociation from heat shock protein 90[J] J Immunol. 2005;174(1):384–394. doi: 10.4049/jimmunol.174.1.384. [DOI] [PubMed] [Google Scholar]

[b15] 15.Bonvini P, Gastaldi T, Falini B, et al. Nucleophosmin-anaplastic lymphoma kinase (NPM-ALK), a novel Hsp90-client tyrosine kinase: down-regulation of NPM-ALK expression and tyrosine phosphorylation in ALK(+) CD30(+) lymphoma cells by the Hsp90 antagonist 17-allylamino,17-demethoxygeldanamycin[J] Cancer Res. 2002;62(5):1559–1566. [PubMed] [Google Scholar]

[b16] 16.Bonvini P, Dalla Rosa H, Vignes N, et al. Ubiquitination and proteasomal degradation of nucleophosmin-anaplastic lymphoma kinase induced by 17-allylamino-demethoxygeldanamycin: role of the co-chaperone carboxyl heat shock protein 70-interacting protein[J] Cancer Res. 2004;64(9):3256–3264. doi: 10.1158/0008-5472.CAN-03-3531. [DOI] [PubMed] [Google Scholar]

[b17] 17.Yao Q, Nishiuchi R, Li Q, et al. FLT3 expressing leukemias are selectively sensitive to inhibitors of the molecular chaperone heat shock protein 90 through destabilization of signal transduction-associated kinases[J] Clin Cancer Res. 2003;9(12):4483–4493. [PubMed] [Google Scholar]

[b18] 18.Beliakoff J, Bagatell R, Paine-Murrieta G, et al. Hormone-refractory breast cancer remains sensitive to the antitumor activity of heat shock protein 90 inhibitors[J] Clin Cancer Res. 2003;9(13):4961–4971. [PubMed] [Google Scholar]

[b19] 19.Solit DB, Chiosis G. Development and application of Hsp90 inhibitors[J] Drug Discov Today. 2008;13(1-2):38–43. doi: 10.1016/j.drudis.2007.10.007. [DOI] [PubMed] [Google Scholar]

[b20] 20.Mahmud SA, Manlove LS, Farrar MA. Interleukin-2 and STAT5 in regulatory T cell development and function[J] JAKSTAT. 2013;2(1):e23154. doi: 10.4161/jkst.23154. [DOI] [PMC free article] [PubMed] [Google Scholar]

[b21] 21.Fumo G, Akin C, Metcalfe DD, et al. 17-Allylamino-17-demethoxygeldanamycin(17-AAG) is effective in down-regulating mutated, constitutively activated KIT protein in human mast cells[J] Blood. 2004;103(3):1078–1084. doi: 10.1182/blood-2003-07-2477. [DOI] [PubMed] [Google Scholar]

[b22] 22.Rao R, Nalluri S, Fiskus W, et al. Heat shock protein 90 inhibition depletes TrkA levels and signaling in human acute leukemia cells[J] Mol Cancer Ther. 2010;9(8):2232–2242. doi: 10.1158/1535-7163.MCT-10-0336. [DOI] [PMC free article] [PubMed] [Google Scholar]

[b23] 23.Schoof N, von Bonin F, Trümper L, et al. HSP90 is essential for Jak-STAT signaling in classical Hodgkin lymphoma cells[J] Cell Commun Signal. 2009;7:17. doi: 10.1186/1478-811X-7-17. [DOI] [PMC free article] [PubMed] [Google Scholar]

[b24] 24.Whitesell L, Lindquist SL. HSP90 and the chaperoning of cancer[J] Nat Rev Cancer. 2005;5(10):761–772. doi: 10.1038/nrc1716. [DOI] [PubMed] [Google Scholar]

[b25] 25.Pearl LH, Prodromou C, Workman P. The Hsp90 molecular chaperone: an open and shut case for treatment[J] Biochem J. 2008;410(3):439–453. doi: 10.1042/BJ20071640. [DOI] [PubMed] [Google Scholar]

[b26] 26.Jez JM, Chen JC, Rastelli G, et al. Crystal structure and molecu-lar modeling of 17-DMAG in complex with human Hsp90[J] Chem Biol. 2003;10(4):361–368. doi: 10.1016/S1074-5521(03)00075-9. [DOI] [PubMed] [Google Scholar]

[b27] 27.Rao R, Lee P, Fiskus W, et al. Co-treatment with heat shock protein 90 inhibitor 17-dimethylaminoethylamino-17-demethoxygeldanamycin (DMAG) and vorinostat: a highly active combination against human mantle cell lymphoma (MCL) cells[J] Cancer Biol Ther. 2009;8(13):1273–1280. doi: 10.4161/cbt.8.13.8726. [DOI] [PMC free article] [PubMed] [Google Scholar]

[b28] 28.Jhaveri K, Modi S. HSP90 inhibitors for cancer therapy and overcoming drug resistance[J] Adv Pharmacol. 2012;65:471–517. doi: 10.1016/B978-0-12-397927-8.00015-4. [DOI] [PubMed] [Google Scholar]

PERMALINK

热休克蛋白90抑制剂17-AAG对HUT-102细胞株JAK3/STAT5信号通路的影响

HSP90 inhibitor 17-AAG plays an important role in JAK3/STAT5 signaling pathways in HTLV-1 infection cell line HUT-102

Qingqing Yang

Huo Tan

Zhiping Fu

Qiang Ma

Jinlong Song

Abstract

目的

方法

结果

结论