Departamento de Farmacologia, Escola Paulista de Medicina (EPM), Universidade Federal de São Paulo (UNIFESP), São Paulo, SP 04044-020, Brazil; Grupo de Fitocomplexos e Sinalização Celular, Escola de Ciências da Saúde, Universidade Anhembi Morumbi, São Paulo, SP 03164-000, Brazil.
Departamento de Farmacologia, Escola Paulista de Medicina (EPM), Universidade Federal de São Paulo (UNIFESP), São Paulo, SP 04044-020, Brazil; Grupo de Fitocomplexos e Sinalização Celular, Escola de Ciências da Saúde, Universidade Anhembi Morumbi, São Paulo, SP 03164-000, Brazil.
J Integr Med. 2019 Mar;17(2):132-140. doi: 10.1016/j.joim.2019.02.001. Epub 2019 Feb 8.
Although Angelica archangelica is a medicinal and aromatic plant with a long history of use for both medicinal and food purposes, there are no studies regarding the antineoplastic activity of its root. This study aimed to evaluate the cytotoxicity and antitumor effects of the crude extract of A. archangelica root (CEAA) on breast cancer.
The cytotoxicity of CEAA against breast adenocarcinoma cells (4T1 and MCF-7) was evaluated by a 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Morphological and biochemical changes were detected by Hoechst 33342/propidium iodide (PI) and annexin V/PI staining. Cytosolic calcium mobilization was evaluated in cells staining with FURA-4NW. Immunoblotting was used to determine the effect of CEAA on anti- and pro-apoptotic proteins (Bcl-2 and Bax, respectively). The 4T1 cell-challenged mice were used for in vivo assay.
Using ultra-high-performance liquid chromatography-mass spectrometry analysis, angelicin, a constituent of the roots and leaves of A. archangelica, was found to be the major constituent of the CEAA evaluated in this study (73 µg/mL). The CEAA was cytotoxic for both breast cancer cell lines studied but not for human fibroblasts. Treatment of 4T1 cells with the CEAA increased Bax protein levels accompanied by decreased Bcl-2 expression, in the presence of cleaved caspase-3 and cytosolic calcium mobilization, suggesting mitochondrial involvement in breast cancer cell death induced by the CEAA in this cell line. No changes on the Bcl-2/Bax ratio were observed in CEAA-treated MCF7 cells. Gavage administration of the CEAA (500 mg/kg) to 4T1 cell-challenged mice significantly decreased tumor growth when compared with untreated animals.
Altogether, our data show the antitumor potential of the CEAA against breast cancer cells in vitro and in vivo. Further research is necessary to better elucidate the pharmacological application of the CEAA in breast cancer therapy.
尽管当归是一种具有悠久药用和食用历史的药用和芳香植物，但目前尚无关于其根的抗肿瘤活性的研究。本研究旨在评估当归根粗提物（CEAA）对乳腺癌的细胞毒性和抗肿瘤作用。
采用 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐（MTT）法评估 CEAA 对乳腺癌腺癌细胞（4T1 和 MCF-7）的细胞毒性。通过 Hoechst 33342/碘化丙啶（PI）和 Annexin V/PI 染色检测形态和生化变化。用 FURA-4NW 染色评估细胞胞质钙动员。用免疫印迹法测定 CEAA 对抗凋亡和促凋亡蛋白（Bcl-2 和 Bax）的影响。用 4T1 细胞攻击的小鼠进行体内试验。
采用超高效液相色谱-质谱分析，发现当归根和叶中的有效成分当归素是本研究评估的 CEAA 的主要成分（73µg/mL）。CEAA 对两种研究的乳腺癌细胞系均具有细胞毒性，但对人成纤维细胞无毒性。CEAA 处理 4T1 细胞可增加 Bax 蛋白水平，同时降低 Bcl-2 表达，伴有裂解的 caspase-3 和胞质钙动员，提示在线粒体参与 CEAA 诱导的乳腺癌细胞死亡。在 CEAA 处理的 MCF7 细胞中，Bcl-2/Bax 比值无变化。与未处理的动物相比，CEAA（500mg/kg）灌胃给药可显著抑制 4T1 细胞攻击的小鼠肿瘤生长。
总之，本研究数据表明 CEAA 具有体外和体内抗肿瘤潜力。需要进一步研究以更好地阐明 CEAA 在乳腺癌治疗中的药理学应用。
