Cardamonin inhibits breast cancer growth by repressing HIF-1α-dependent metabolic reprogramming
Cardamonin inhibits breast cancer growth by repressing HIF-1α-dependent metabolic reprogramming
复制标题
小豆蔻宁通过抑制 HIF-1α 依赖性代谢重编程来抑制乳腺癌生长
DOI：
10.1186/s13046-019-1351-4
复制
发表时间：
2019-08-27
影响因子：
11.3
通讯作者：
Wu, Xiaojun
中科院分区：
文献类型：
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作者：
Jin, Jinmei;Qiu, Shuiping;Wu, Xiaojun
背景小豆蔻甙是从高良姜中分离出来的查耳酮，具有抗炎和抗肿瘤活性。然而，小豆蔻苷抑制乳腺癌进展的分子机制在很大程度上仍有待确定。方法采用CCK-8和Hoechst 33258染色分别检测细胞生长和凋亡。通过荧光素酶报告基因测定来测量 HIF-1α 驱动的转录。使用2-NBDG和L-乳酸测定试剂盒检测葡萄糖摄取和乳酸含量。细胞代谢测定在安捷伦 Seahorse Bioscience XF96 细胞外通量分析仪上进行。用JC-1探针测量线粒体膜电位。 DCFH-DA用于测量ROS水平。通过蛋白质印迹法检测蛋白表达。采用免疫组织化学方法检测肿瘤组织中 HIF-1α、LDHA 和 CD31 的表达。结果小豆蔻苷通过抑制 HIF-1α 介导的细胞代谢，在体外和体内抑制三阴性乳腺癌细胞系 MDA-MB-231 的生长。小豆蔻宁通过抑制 mTOR/p70S6K 通路，在 mRNA 和蛋白质水平抑制 HIF-1α 的表达，随后增强线粒体氧化磷酸化并诱导活性氧 (ROS) 积累。我们还发现小豆蔻苷抑制 Nrf2 依赖性 ROS 清除系统，从而进一步增加细胞内 ROS 水平。最终，细胞内活性氧的积累诱导乳腺癌细胞凋亡。此外，小豆蔻苷治疗减少了葡萄糖的摄取以及乳酸的产生和流出，表明其具有抑制糖酵解过程的功能。结论这些结果揭示了小豆蔻苷在调节癌细胞代谢和抑制乳腺癌进展方面的新功能，并表明其在乳腺癌治疗中的潜力。
BackgroundCardamonin, a chalcone isolated fromAlpiniae katsumadai, has anti-inflammatory and anti-tumor activities. However, the molecular mechanism by which cardamonin inhibits breast cancer progression largely remains to be determined.MethodsCCK-8 and Hoechst 33258 staining were used to detect cell growth and apoptosis, respectively. HIF-1α driven transcription was measured by luciferase reporter assay. Glucose uptake and lactate content were detected with 2-NBDG and L-Lactate Assay Kit. Cell metabolism assays were performed on Agilent’s Seahorse Bioscience XF96 Extracellular Flux Analyzer. Mitochondrial membrane potential was measured with JC-1 probe. DCFH-DA was used to measure ROS level. Protein expression was detected by western blotting assay. Immunohistochemistry was performed to measure the expression of HIF-1α, LDHA and CD31 in tumor tissues.ResultsCardamonin inhibited growth of the triple negative breast cancer cell line MDA-MB-231 in vitro and in vivo by suppressing HIF-1α mediated cell metabolism. Cardamonin inhibited the expression of HIF-1α at mRNA and protein levels by repressing the mTOR/p70S6K pathway, and subsequently enhanced mitochondrial oxidative phosphorylation and induced reactive oxygen species (ROS) accumulation. We also found that cardamonin inhibited the Nrf2-dependent ROS scavenging system which further increased intracellular ROS levels. Eventually, accumulation of the intracellular ROS induced apoptosis in breast cancer cells. In addition, cardamonin treatment reduced glucose uptake as well as lactic acid production and efflux, suggesting its function in repressing the glycolysis process.ConclusionsThese results reveal novel function of cardamonin in modulating cancer cell metabolism and suppressing breast cancer progression, and suggest its potential for breast cancer treatment.