Oroxylin A promotes PTEN-mediated negative regulation of MDM2 transcription via SIRT3-mediated deacetylation to stabilize p53 and inhibit glycolysis in wt-p53 cancer cells.
Oroxylin A promotes PTEN-mediated negative regulation of MDM2 transcription via SIRT3-mediated deacetylation to stabilize p53 and inhibit glycolysis in wt-p53 cancer cells.
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Oroxylin A 通过 SIRT3 介导的脱乙酰化促进 PTEN 介导的 MDM2 转录负调节，以稳定 p53 并抑制 wt-p53 癌细胞中的糖酵解
DOI：
10.1186/s13045-015-0137-1
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发表时间：
2015-04-23
影响因子：
28.5
通讯作者：
Wei L
中科院分区：
文献类型：
--
作者：
Zhao K;Zhou Y;Qiao C;Ni T;Li Z;Wang X;Guo Q;Lu N;Wei L
p53在调节癌症代谢重编程中起重要作用，如有氧糖酵解。oroxlin A是一种天然活性类黄酮，体外和体内均有很强的抗癌作用。方法采用swt-p53 （MCF-7和HCT116细胞）癌细胞和p53-null H1299癌细胞。采用乳酸生成检测试剂盒和Amplex红葡萄糖检测试剂盒分析葡萄糖摄取和乳酸生成。然后，分别采用Western blotting和定量聚合酶链反应（PCR）技术对p53、小鼠双分钟2 （MDM2）和p53靶向糖酵解酶的蛋白水平和RNA水平进行定量分析。采用免疫沉淀法评估p53、MDM2和sirtun -3 （SIRT3）之间的结合，以及磷酸酶和紧张素同源物（PTEN）的去乙酰化。采用报告试验来评估PTEN的转录活性。在体内，研究了oroxylin A对裸鼠移植瘤接种MCF-7或HCT116细胞的作用。结果我们分析了oroxylin A调控wt-p53癌细胞中p53水平和糖酵解代谢的潜在机制，发现oroxylin A通过上调p53水平抑制糖酵解。Oroxylin A不直接影响wt-p53的转录，而是通过下调wt-p53癌细胞中MDM2的转录抑制MDM2介导的p53降解。在进一步的研究中，我们发现oroxylin A通过促进磷酸酶和紧张素同源物的脂质磷酸酶活性来诱导MDM2转录的减少，这种活性通过sirtuin3介导的去乙酰化而上调。在体内，oroxylin A抑制裸鼠接种MCF-7或HCT116细胞的肿瘤生长。oroxylin A也能下调肿瘤组织中MDM2蛋白的表达。结论这些结果提供了一个不依赖于p53的MDM2转录机制，揭示了oroxylin a在wt-p53和mutp53癌细胞中糖酵解调节的潜力。本研究对奥罗霉素A的抗癌作用的研究具有重要意义，并为奥罗霉素A在癌症患者中的临床试验提供了理论依据。
Introductionp53 plays important roles in regulating the metabolic reprogramming of cancer, such as aerobic glycolysis. Oroxylin A is a natural active flavonoid with strong anticancer effects bothin vitroandin vivo.Methodswt-p53 (MCF-7 and HCT116 cells) cancer cells and p53-null H1299 cancer cells were used. The glucose uptake and lactate production were analyzed using Lactic Acid production Detection kit and the Amplex Red Glucose Assay Kit. Then, the protein levels and RNA levels of p53, mouse double minute 2 (MDM2), and p53-targeted glycolytic enzymes were quantified using Western blotting and quantitative polymerase chain reaction (PCR), respectively. Immunoprecipitation were performed to assess the binding between p53, MDM2, and sirtuin-3 (SIRT3), and the deacetylation of phosphatase and tensin homolog (PTEN). Reporter assays were performed to assess the transcriptional activity of PTEN.In vivo, effects of oroxylin A was investigated in nude mice xenograft tumor-inoculated MCF-7 or HCT116 cells.ResultsHere, we analyzed the underlying mechanisms that oroxylin A regulated p53 level and glycolytic metabolism in wt-p53 cancer cells, and found that oroxylin A inhibited glycolysis through upregulating p53 level. Oroxylin A did not directly affect the transcription of wt-p53, but suppressed the MDM2-mediated degradation of p53 via downregulating MDM2 transcription in wt-p53 cancer cells. In further studies, we found that oroxylin A induced a reduction in MDM2 transcription by promoting the lipid phosphatase activity of phosphatase and tensin homolog, which was upregulated via sirtuin3-mediated deacetylation.In vivo, oroxylin A inhibited the tumor growth of nude mice-inoculated MCF-7 or HCT116 cells. The expression of MDM2 protein in tumor tissue was downregulated by oroxylin A as well.ConclusionsThese results provide a p53-independent mechanism of MDM2 transcription and reveal the potential of oroxylin A on glycolytic regulation in both wt-p53 and mut-p53 cancer cells. The studies have important implications for the investigation on anticancer effects of oroxylin A, and provide the academic basis for the clinical trial of oroxylin A in cancer patients.
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影响因子：
10.9
作者：
Ortega-Molina, Ana;Serrano, Manuel
通讯作者：
Serrano, Manuel
Oroxylin A 通过阻断血管内皮生长因子诱导的 KDR/Flk-1 磷酸化来抑制血管生成
DOI：
10.1007/s00432-009-0705-2
发表时间：
2010-05-01
影响因子：
3.6
作者：
Gao, Ying;Lu, Na;Guo, Qinglong
通讯作者：
Guo, Qinglong
影响因子：
5.3
作者：
Iwahara, Toshinori;Bonasio, Roberto;Reinberg, Danny
通讯作者：
Reinberg, Danny
DOI：
10.1073/pnas.95.23.13513
发表时间：
1998-11-10
影响因子：
11.1
作者：
Myers, MP;Pass, I;Tonks, NK
通讯作者：
Tonks, NK
影响因子：
4.8
作者：
Chang, CJ;Freeman, DJ;Wu, H
通讯作者：
Wu, H