摘要
Marsdeniae tenacissimae extract(MTE), commonly known as Xiao-Ai-Ping in China, is a traditional Chinese herb medicine capable of inhibiting proliferation and metastasis and boosting apoptosis in various cancer cells. However, little is known about the contribution of MTE towards tumor angiogenesis and the underlying mechanism. The present study aimed to evaluate the effects of MTE on the proliferation and apoptosis of human umbilical vein endothelial cells(HUVECs) and the molecular ism. 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazolium, inner salt(MTS) and PI-stained flow cytometry assays revealed that MTE dose-dependently reduced the proliferation of HUVECs by arresting cell cycle at S phase(P < 0.05). Annexin V-FITC/PI-stained flow cytometry confirmed that MTE(160 μL·m L-1) enhanced the apoptosis of HUVECs significantly(P < 0.001). Real-time quantitative RT-PCR and Western blot analyses showed an increase in Bax expression and a sharply decline in Bcl-2 expression; caspase-3 was activated simultaneously in a dose-dependent manner(P < 0.05). Further study observed the dose-dependent down-regulation of vascular endothelial growth factor(VEGF) receptor-2(VEGFR-2), P2Y6 receptor(P2Y6R), and chemokine(C-C motif) ligand 2(CCL-2), along with the activation of PKC δ and up-regulation of p53 in a dose-dependent manner in MTE-treated selected cells(P < 0.05). Collectively, the results from the present study suggested that MTE suppressed the proliferation by attenuating CCL-2-mediated VEGF/VEGFR2 interactions and promoted the apoptosis through PKCδ-induced p53-dependent mitochondrial pathway in HUVECs, supporting that MTE may be developed as a potent anti-cancer medicine.
Marsdeniae tenacissimae extract（MTE）, commonly known as Xiao-Ai-Ping in China, is a traditional Chinese herb medicine capable of inhibiting proliferation and metastasis and boosting apoptosis in various cancer cells. However, little is known about the contribution of MTE towards tumor angiogenesis and the underlying mechanism. The present study aimed to evaluate the effects of MTE on the proliferation and apoptosis of human umbilical vein endothelial cells（HUVECs） and the molecular ism. 3-（4,5-dimethylthiazol-2-yl）-5（3-carboxymethoxyphenyl）-2-（4-sulfopheny）-2H-tetrazolium, inner salt（MTS） and PI-stained flow cytometry assays revealed that MTE dose-dependently reduced the proliferation of HUVECs by arresting cell cycle at S phase（P 〈 0.05）. Annexin V-FITC/PI-stained flow cytometry confirmed that MTE（160 μL·m L-1） enhanced the apoptosis of HUVECs significantly（P 〈 0.001）. Real-time quantitative RT-PCR and Western blot analyses showed an increase in Bax expression and a sharply decline in Bcl-2 expression; caspase-3 was activated simultaneously in a dose-dependent manner（P 〈 0.05）. Further study observed the dose-dependent down-regulation of vascular endothelial growth factor（VEGF） receptor-2（VEGFR-2）, P2Y6 receptor（P2Y6R）, and chemokine（C-C motif） ligand 2（CCL-2）, along with the activation of PKC δ and up-regulation of p53 in a dose-dependent manner in MTE-treated selected cells（P 〈 0.05）. Collectively, the results from the present study suggested that MTE suppressed the proliferation by attenuating CCL-2-mediated VEGF/VEGFR2 interactions and promoted the apoptosis through PKCδ-induced p53-dependent mitochondrial pathway in HUVECs, supporting that MTE may be developed as a potent anti-cancer medicine.
基金
supported by the National Natural Science Foundation of China(No.81501824)
the Natural Science Foundation of Zhejiang Province(Nos.Y15H280010 and LY15C090004)
the Analysis and Measurement Foundation of Zhejiang Province(No.2015C37001)
the Key Project of Chinese medicine in Zhejiang Province(No.2015ZZ001)
the Traditional Chinese Medicine Scientific Research and Outstanding Young Talent Foundation of Zhejiang Province(Nos.2014ZB007 and 2014ZQ005)
the Medicine and Health Research Foundation of Zhejiang Province(Nos.2015ZDA002,2015KYA028,2014KYA233,2015KYB021,and 2017179748)
