Effects of AG490 and S3I-201 on regulation of the JAK/STAT3 signaling pathway in relation to angiogenesis in TRAIL-resistant prostate cancer cells in vitro.
Effects of AG490 and S3I-201 on regulation of the JAK/STAT3 signaling pathway in relation to angiogenesis in TRAIL-resistant prostate cancer cells in vitro.
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DOI：
10.3892/ol.2014.1795
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发表时间：
2014-03
期刊：
影响因子：
2.9
通讯作者：
Sozen S
中科院分区：
文献类型：
--
作者：
Gurbuz V;Konac E;Varol N;Yilmaz A;Gurocak S;Menevse S;Sozen S
本研究的目的是分析阻断信号通路的分子机制及其对体外前列腺癌(CAP)细胞进展的影响。用白细胞介素6(IL-6)刺激LNCaP人CAP细胞系，同时加入JAK 2(AG490)、信号转导和转录激活剂3[(STAT3)S3I-201]抑制剂和肿瘤坏死因子相关的凋亡诱导配体(TRAIL)。检测细胞毒活性、磷酸化(P)-STAT3蛋白活性、蛋白水平半胱氨酸天冬氨酸氨基转移酶(CASP)3活性、血管内皮生长因子(VEGF)A、VEGFC、血管内皮生长因子受体2、STAT3、基质金属蛋白酶-2、髓系细胞白血病序列1(MCL-1)、CASP8和CASP9信使RNA(MRNA)水平。DAPI染色和末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)法检测细胞形态和细胞凋亡。IL-6以剂量和时间依赖的方式快速诱导STAT3的磷酸化，在浓度为25 ng/ml的3h达到高峰，50μM的AG490和300μM的S3I-201抑制STAT3的激活。Western blotting结果显示，经AG490和S3I-201处理后，LNCaP细胞中p-STAT3蛋白的表达显著降低。AG490和S3I-201诱导VEGFA、MCL-1和STAT3基因表达下调，CASP8和CASP9基因转录水平上调。此外，这些抑制剂还增加了CASP3蛋白的水平。AG490-和S3I-201-TRAIL的组合并没有导致这种效应的增加。DAPI染色和TUNEL法检测结果基本一致。据我们所知，这是第一次研究AG490或S3I-201可能的临床应用，以及减少使用高细胞毒性的化疗药物，以发挥针对JAK/STAT3途径的凋亡效应。
The aim of the present study was to analyze the molecular mechanisms involved in blocking the signaling pathway and the effects of this on the progression of prostate cancer (CaP) cells in vitro. LNCaP human CaP cell line was stimulated with interleukin-6 (IL-6) in the presence/absence of Janus kinase (JAK) 2 (AG490), signal transducer and activator of transcription 3 [(STAT3) S3I-201] inhibitors and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Cytotoxic activity, the activation of phosphorylated (p)-STAT3 protein, caspase (CASP) 3 activity at protein level, vascular endothelial growth factor (VEGF) A, VEGFC, vascular endothelial growth factor receptor 2, STAT3, matrix metalloproteinase-2, myeloid cell leukemia sequence 1 (MCL-1), CASP8 and CASP9 messenger RNA (mRNA) levels were determined. Morphology and apoptosis were confirmed by DAPI staining and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. IL-6 rapidly induced the phosphorylation of STAT3 in a dose- and time-dependent manner with a peak expression at 3 h at a concentration of 25 ng/ml. In addition, AG490 (50 μM) and S3I-201 (300 μM) inhibited STAT3 activation. Western blotting results revealed that p-STAT3 protein expression decreased significantly with AG490 and S3I-201 treatment in LNCaP cells. AG490 and S3I-201 induced the downregulation of VEGFA, MCL-1 and STAT3 and the upregulation of CASP8 and CASP9 mRNA transcription levels. In addition, the inhibitors increased the level of CASP3 protein. Combinations of AG490- and S3I-201-TRAIL did not result in an increase in this effect. Parallel results were found by DAPI staining and TUNEL assay. To the best of our knowledge, this is the first study to investigate the possible clinical use of AG490 or S3I-201, together with the reduced use of chemotherapeutic agents with high cytotoxicity, for their ability to exert an apoptotic effect, targeting the JAK/STAT3 pathway.
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影响因子：
8
作者：
Lin, L.;Amin, R.;Gallicano, G. I.;Glasgow, E.;Jogunoori, W.;Jessup, J. M.;Zasloff, M.;Marshall, J. L.;Shetty, K.;Johnson, L.;Mishra, L.;He, A. R.
通讯作者：
He, A. R.
影响因子：
15.9
作者：
Epling-Burnette, PK;Liu, JH;Loughran, TP
通讯作者：
Loughran, TP
DOI：
10.1186/1756-9966-29-51
发表时间：
2010-05-19
期刊：
Journal of experimental & clinical cancer research : CR
影响因子：
--
作者：
Huang C;Yang G;Jiang T;Huang K;Cao J;Qiu Z
通讯作者：
Qiu Z
影响因子：
6.7
作者：
Chen, Wei;Shen, Xuning;Liang, Tingbo
通讯作者：
Liang, Tingbo
影响因子：
44.1
作者：
Lanuti, Paola;Bertagnolo, Valeria;Marchisio, Marco
通讯作者：
Marchisio, Marco