Molecular and cellular pharmacology
Quercetin inhibits angiogenesis by targeting calcineurin in the xenograft model of human breast cancer
Introduction
Breast cancer is the most common malignant tumor in women (DeSantis et al., 2014). Angiogenesis plays an essential role in the development, invasion, and metastasis of breast cancer (Schneider and Miller, 2005). Developing novel anti-angiogenic agents is an important strategy for treating patients in clinic.
Vascular endothelial growth factor receptor 2 (VEGFR2) mediated calcineurin/nuclear factor of activated T-cells (NFAT) pathway plays leading roles in the angiogenesis of breast cancer (Hoeben et al., 2004). Calcineurin is a target enzyme in this pathway (Mancini and Toker, 2009). In endothelia, VEGFR2 activates calcineurin, triggers NFAT translocated into nucleus, and up-regulates angiogenic factors (Muller and Rao, 2010). Tacrolimus (FK506) is a calcineurin inhibitor, which blocks calcineurin/NFAT pathway, and consequently suppresses angiogenesis (Mancini and Toker, 2009, Muller and Rao, 2010). FK506 also inhibits breast tumor growth in vivo, and reduces vessel formation of human umbilical vein endothelial cells (HUVEC) in vitro (Siamakpour-Reihani et al., 2011). However, FK506 has not been used in clinic for cancer owing to its immunosuppressive effect, so the novel inhibitors targeting calcineurin with minor adverse effect might provide a major advantage in treating breast cancer.
Quercetin (Qu), a flavonoid abundant in fruits, plants, and vegetables, is characterized by a phenyl benzo(c)pyrone-derived structure (Nabavi et al., 2015). It is present in plants in many different glycosidic forms, with the most widespread form of quercetin-3-rutinoside (Erlund, 2004). It has exhibited antioxidant, cardioprotective (Nabavi et al., 2012b, Nabavi et al., 2012a), vasodilatory (Larson et al., 2012), live-protective (Nabavi et al., 2012a, Nabavi et al., 2012b), anti-inflammatory, and anti-cancer activities (Nabavi et al., 2015), with almost no human toxicity. Its anti-cancer action is associated with anti-proliferation, pro-apoptosis and anti-angiogenesis (Cheng et al., 2010, Chirumbolo, 2013). Qu induces breast cancer cell apoptosis via trigging extrinsic or mitochondria-related apoptotic pathways (de Oliveira et al., 2015). It interacts with the intracellular signaling cascades for cancer cell survival or proliferation, such as PI3K/AKT and Ras Raf/MEK/ERK1/2/RSK signaling pathways (Dajas, 2012). Qu can also eliminate damaged DNA, inhibit cancer associated inflammatory and obesity pathways (Burd, 2011).
In addition, previous studies have shown that Qu reduces serum VEGF levels of tumor bearing animals (Camargo et al., 2011) and inhibits VEGF secretion of human breast cancer MCF-7 cells (Oh et al., 2010). And in vitro observations find that Qu exerts a strong inhibitory effect on calcineurin (Lei et al., 2011). Therefore, in the present study, we hypothesize that calcineurin is the key target of Qu in the tumor angiogenesis. Anti-tumor and anti-angiogenesis effects of Qu were assessed in a human breast cancer xenograft model in nude mice. Furthermore, the underlying mechanisms of anti-angiogenic activities of Qu were demonstrated as calcineurin activity, endothelial NFAT activation, subcellular location and gene expression of molecules associated in VEGFR2 mediated calcineurin/NFAT pathway for angiogenesis.
Section snippets
Animals
Thirty two female BALB/c nude mice were obtained from Animal Centre of the Chinese Academy of Medical Sciences. All animals were kept under a 12-h-light/-dark cycle, temperature (25.0±0.2 °C) and humidity (45±2%) controlled specific pathogen free environment. The animals were acclimatized for one week, fed standard rodent pellets and allowed free access to filtered water. All experimental procedures were approved by the Ethics Committee of the Institute of Medicinal Plant Development, CAMS &
Quercetin inhibits MCF-7 breast cancer xenograft growth in mice
The effect of Qu on the growth of xenograft tumor in nude mice was evaluated. Volume ratios of tumors were the largest in model animals, and these were significantly decreased in TAM (P<0.01), FK506 (P<0.05) and Qu (P<0.01) treated animals. Similarly, the weight ratio of tumors in each treatment group is significantly decreased than that in model group. The inhibitory rates of tumor growth in TAM, FK506 and Qu group were 22.86%, 14.05% and 18.26%, respectively (Table 1).
Quercetin induces tumor necrosis
H&E staining showed
Discussion
Angiogenesis plays a critical role in the development of breast cancer (Schneider and Miller, 2005). Calcineurin/NFAT pathway regulated by VEGFR2 is highly activated in human breast tumor endothelia (Courtwright et al., 2009, Siamakpour-Reihani et al., 2011). NFAT is a transcription factor that mediates vascular development and angiogenesis, and calcineurin is the key enzyme inducing NFAT activation (Mancini and Toker, 2009). FK506 binds to immunophilin FKBP12 and inhibits calcineurin
Conclusions
In conclusion, this study demonstrated the antitumor and antiangiogenic effect of Qu on breast cancer in vivo. It indicated that Qu inhibited breast cancer angiogenesis, at least in part via targeting calcineurin/NFAT pathway. Therefore, this study provides for the further study of Qu as a novel antiangiogenic agent for the treatment of cancers. And other potential mechanisms of Qu on molecular links between inflammation and tumor angiogenesis might be identified. Furthermore, the most
Declaration of interest
The authors have declared no conflict of interest.
Acknowledgements
This work was supported by the Beijing Natural Science Foundation (Grant no. 7132150), Education Ministry Science Foundation of the People's Republic of China (Grant no. 108019), and National Natural Science Foundation of China (Grant no. 81541082).
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These authors contributed equally to this work.