NF-κB and pSTAT3 synergistically drive G6PD overexpression and facilitate sensitivity to G6PD inhibition in ccRCC.
NF-κB and pSTAT3 synergistically drive G6PD overexpression and facilitate sensitivity to G6PD inhibition in ccRCC.
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NF-κB 和 pSTAT3 协同驱动 G6PD 过度表达并促进 ccRCC 中对 G6PD 抑制的敏感性
DOI：
10.1186/s12935-020-01576-2
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发表时间：
2020
影响因子：
5.8
通讯作者：
Zhu Y
中科院分区：
文献类型：
--
作者：
Zhang Q;Yang Z;Ni Y;Bai H;Han Q;Yi Z;Yi X;Agbana YL;Kuang Y;Zhu Y
研究背景葡萄糖6-磷酸脱氢酶（G6 PD）在肿瘤细胞代谢重编程中起重要作用，并参与某些肿瘤的发生。来自我们实验室的先前结果表明，过表达的G6 PD是透明细胞肾细胞癌（ccRCC）（肾癌的最常见亚型）中的潜在预后生物标志物。G6 PD可通过促进活性氧（ROS）-磷酸化信号转导子和转录激活子3（pSTAT 3）的激活和促进ROS-MAPK-MMP 2轴途径促进ccRCC的生长和侵袭。方法采用实时荧光定量RT-PCR和Western blot方法，检测活性氧刺激剂或清除剂处理ccRCC细胞后，细胞内活性氧积累对NF-κB信号通路和G6 PD表达的影响。采用实时荧光定量RT-PCR、Western blot、荧光素酶和ChIP等方法，分析NF-κ B信号通路对肾癌细胞G6 PD转录的调控作用。分别进行ChIP和Co-IP测定以证明NF-κB和pSTAT 3的蛋白-DNA和蛋白-蛋白相互作用。结果ROS诱导的NF-κB和pSTAT 3信号通路的过度激活可相互激活，并在G6 PD异常转录调控中表现出交叉作用。其机制是NF-κB信号通路通过与p65而不是p50的直接DNA-蛋白质相互作用促进G6 PD的转录。p65和pSTAT 3形成p65/pSTAT 3复合物，占据G6 PD启动子上的pSTAT 3结合位点，并在易化的G6 PD过表达后促进ccRCC增殖。在ccRCC组织中，G6 PD、pSTAT 3和p65高度表达并且彼此正相关，证实NF-κB和pSTAT 3协同促进G6 PD过表达。结论ROS诱导的NF-κB和pSTAT 3信号通路的异常通过形成p65/pSTAT 3复合物协同驱动G6 PD的转录。此外，G6 PD活性抑制可能是用于ccRCC治疗的有希望的治疗策略。
BackgroundGlucose 6-phosphate dehydrogenase (G6PD) serves key roles in cancer cell metabolic reprogramming, and has been reported to be involved in certain carcinogenesis. Previous results from our laboratory demonstrated that overexpressed G6PD was a potential prognostic biomarker in clear cell renal cell carcinoma (ccRCC), the most common subtype of kidney cancer. G6PD could stimulate ccRCC growth and invasion through facilitating reactive oxygen species (ROS)-phosphorylated signal transducer and activator of transcription 3 (pSTAT3) activation and ROS-MAPK-MMP2 axis pathway, respectively. However, the reasons for ectopic G6PD overexpression and the proliferation repressive effect of G6PD inhibition in ccRCC are still unclear.MethodsThe impact of ROS accumulation on NF-κB signaling pathway and G6PD expression was determined by real-time RT-PCR and Western blot in ccRCC cells following treatment with ROS stimulator or scavenger. The regulatory function of NF-κB signaling pathway in G6PD transcription was analyzed by real-time RT-PCR, Western blot, luciferase and ChIP assay in ccRCC cells following treatment with NF-κB signaling activator/inhibitor or lentivirus infection. ChIP and Co-IP assay was performed to demonstrate protein-DNA and protein–protein interaction of NF-κB and pSTAT3, respectively. MTS assay, human tissue detection and xenograft model were conducted to characterize the association between NF-κB, pSTAT3, G6PD expression level and proliferation functions.ResultsROS-stimulated NF-κB and pSTAT3 signaling over-activation could activate each other, and exhibit cross-talks in G6PD aberrant transcriptional regulation. The underlying mechanism was that NF-κB signaling pathway facilitated G6PD transcription via direct DNA–protein interaction with p65 instead of p50. p65 and pSTAT3 formed a p65/pSTAT3 complex, occupied the pSTAT3-binding site on G6PD promoter, and contributed to ccRCC proliferation following facilitated G6PD overexpression. G6PD, pSTAT3, and p65 were highly expressed and positively correlated with each other in ccRCC tissues, confirming that NF-κB and pSTAT3 synergistically promote G6PD overexpression. Moreover, G6PD inhibitor exhibited tumor-suppressor activities in ccRCC and attenuated the growth of ccRCC cells both in vitro and in vivo.ConclusionROS-stimulated aberrations of NF-κB and pSTAT3 signaling pathway synergistically drive G6PD transcription through forming a p65/pSTAT3 complex. Moreover, G6PD activity inhibition may be a promising therapeutic strategy for ccRCC treatment.
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DOI：
10.1038/nrurol.2010.47
发表时间：
2010-05
期刊：
Nature reviews. Urology
影响因子：
--
作者：
通讯作者：
--
二氢杨梅素通过 ROS-STAT3 信号促进头颈鳞状细胞癌的自噬和凋亡
影响因子：
--
作者：
Fan TF;Wu TF;Bu LL;Ma SR;Li YC;Mao L;Sun ZJ;Zhang WF
通讯作者：
Zhang WF
DOI：
10.3233/kca-170015
发表时间：
2017-11-27
期刊：
Kidney cancer (Clifton, Va.)
影响因子：
--
作者：
Ghatalia P;Koenigsberg R;Pisarcik D;Handorf EA;Geynisman DM;Zibelman M
通讯作者：
Zibelman M
影响因子：
12.8
作者：
Kim SJ;Yoon S
通讯作者：
Yoon S
影响因子：
4
作者：
Gionet, Nathalie;Jansson, Deidre;Pratt, M. A. Christine
通讯作者：
Pratt, M. A. Christine