Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100193, China; Department of Oncology of Integrative Chinese and Western Medicine, China-Japan Friendship Hospital, Beijing 100029, China.
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100193, China.
J Ethnopharmacol. 2018 Aug 10;222:261-269. doi: 10.1016/j.jep.2018.01.013. Epub 2018 Jan 11.
Qingdu granule (QDG), a traditional Chinese herbal prescription, had anti-tumor effect on breast cancer. However the underlying mechanism of QDG was unclear.
The present study aimed to investigate whether QDG could inhibit angiogenesis of breast cancer via acting on nuclear factor of activated T-cells (NFAT) signaling pathway. This was implicated in human umbilical vein endothelial cells (HUVECs) in vitro and breast cancer xenograft model in vivo.
The VEGF (15.58 ng/mL) induced human umbilical vein endothelial cells (HUVECs) were treated with serum samples containing tamoxifen (TAM), tacrolimus (FK506), or QDG with three dosages. The migration and canalization capacities of HUVECs were evaluated by transwell migration and tube formation assay. In 72 h-cultured HUVECs, The gene expression, protein amount, and nuclear translocation of NFATc3 were measured. The anti-tumor and anti-angiogenic effects of QDG in vivo were investigated in breast cancer xenograft model. The serum VEGF levels, microvessel density, and protein expressions (immunohistochemistry and western blot) of VEGF, VEGFR2 and NFATc3 were detected.
The results showed that, QDG significantly inhibited HUVEC migration and tube formation. It downregulated NFATc3 gene expression, decreased NFATc3 protein amount, and reduced the ratio of NFATc3 nuclear translocation in HUVECs. In breast cancer xenograft model, QDG treatment significantly suppressed tumor growth, inhibited VEGF release, and decreased microvessel density. QDG reduced protein expressions of VEGF, VEGFR2 and NFATc3.
The results suggested that QDG showed anti-angiogenic effects of breast cancer both in vitro and in vivo. The mechanism might be partially associated with inhibiting NFAT signaling pathway.
清毒颗粒（QDG）是一种中药复方，对乳腺癌具有抗肿瘤作用。然而，其作用机制尚不清楚。
本研究旨在探讨 QDG 是否通过核因子活化 T 细胞（NFAT）信号通路抑制乳腺癌血管生成。这在体外人脐静脉内皮细胞（HUVEC）和体内乳腺癌异种移植模型中得到了证实。
用含有他莫昔芬（TAM）、他克莫司（FK506）或 QDG 三种剂量的血清样品处理 VEGF（15.58ng/mL）诱导的 HUVEC。通过 Transwell 迁移和管形成实验评估 HUVEC 的迁移和管形成能力。在 72h 培养的 HUVEC 中，测量 NFATc3 的基因表达、蛋白含量和核转位。在乳腺癌异种移植模型中研究 QDG 的体内抗肿瘤和抗血管生成作用。检测血清 VEGF 水平、微血管密度以及 VEGF、VEGFR2 和 NFATc3 的蛋白表达（免疫组织化学和 Western blot）。
结果表明，QDG 显著抑制 HUVEC 迁移和管形成。它下调 NFATc3 基因表达，降低 NFATc3 蛋白含量，并减少 HUVEC 中 NFATc3 核转位的比例。在乳腺癌异种移植模型中，QDG 治疗显著抑制肿瘤生长，抑制 VEGF 释放，降低微血管密度。QDG 降低了 VEGF、VEGFR2 和 NFATc3 的蛋白表达。
结果表明，QDG 在体内外均表现出抗乳腺癌血管生成作用。其机制部分与抑制 NFAT 信号通路有关。
