Lee Eun-Ok, Lee Hyo-Jung, Hwang Hwa-Soo, Ahn Kyoo-Seok, Chae Chanhee, Kang Kyung-Sun, Lu Junxuan, Kim Sung-Hoon
Graduate School of East-West Medical Science, Kyunghee University, Kiheungeup, Yongin 449-701, Republic of Korea.
Carcinogenesis. 2006 Oct;27(10):2059-69. doi: 10.1093/carcin/bgl055. Epub 2006 May 4.
Vitis amurensis Rupr. (Vitaceae) has long been used in Chinese/Oriental herbal medicine for the treatment of cancer, but its active compounds and mechanisms of action have not been well studied. To this end, we isolated from its root heyneanol A (HA), which is a tetramer of resveratrol (RES), and established the in vivo antitumor activity of HA using the mouse Lewis lung carcinoma (LLC) model. We administered HA and RES by daily intraperitonial injection to C57BL/6 mice that were subcutaneously inoculated with LLC cells. HA dose-dependently decreased tumor growth without any adverse effect on body weight and seemed more potent than RES. The tumor inhibitory effects were accompanied by a marked increase in tumor cell apoptosis detected by cleaved caspase-3 and TUNEL assays and decreased tumor cell proliferation index and tumor microvessel density, supporting the involvement of apoptotic and anti-angiogenic activities in the anticancer effects. We next investigated the cellular and molecular processes that mediate the apoptosis and anti-angiogenesis effects using cell culture models. Mechanistically, treatment of LLC cells in vitro with HA or RES significantly increased apoptotic cells. Both HA- and RES-induced cleavage of caspase-9 and caspase-3 and PARP were completely blocked by a pan caspase inhibitor, Z-VAD-FMK. In addition, HA and RES suppressed the basic fibroblast growth factor (bFGF)-induced proliferation and capillary differentiation of human umbilical vein endothelial cells, and inhibited the binding of bFGF to its receptor in a test tube assay and the bFGF-induced vascularization of Matrigel plugs in vivo. Remarkably, HA was fairly stable in cell culture medium and did not undergo intracellular conversion to RES. Therefore, HA is an active anticancer compound that induces caspase-mediated cancer cell apoptosis and inhibits angiogenesis rivaling the potency of RES and merits further evaluation for cancer chemoprevention.
山葡萄（葡萄科）长期以来在中国/东方草药中用于治疗癌症，但其活性成分及作用机制尚未得到充分研究。为此，我们从其根部分离出了白藜芦醇A（HA），它是白藜芦醇（RES）的四聚体，并使用小鼠Lewis肺癌（LLC）模型确定了HA的体内抗肿瘤活性。我们通过每日腹腔注射将HA和RES给予皮下接种LLC细胞的C57BL/6小鼠。HA剂量依赖性地降低肿瘤生长，对体重没有任何不良影响，并且似乎比RES更有效。肿瘤抑制作用伴随着通过裂解的半胱天冬酶-3和TUNEL检测法检测到的肿瘤细胞凋亡显著增加，以及肿瘤细胞增殖指数和肿瘤微血管密度降低，这支持了凋亡和抗血管生成活性参与抗癌作用。接下来，我们使用细胞培养模型研究了介导凋亡和抗血管生成作用的细胞和分子过程。从机制上讲，用HA或RES体外处理LLC细胞可显著增加凋亡细胞。HA和RES诱导的半胱天冬酶-9、半胱天冬酶-3和PARP的裂解均被泛半胱天冬酶抑制剂Z-VAD-FMK完全阻断。此外，HA和RES抑制碱性成纤维细胞生长因子（bFGF）诱导的人脐静脉内皮细胞增殖和毛细血管分化，并在试管试验中抑制bFGF与其受体的结合以及bFGF诱导的体内基质胶塞血管生成。值得注意的是，HA在细胞培养基中相当稳定，不会在细胞内转化为RES。因此，HA是一种活性抗癌化合物，可诱导半胱天冬酶介导的癌细胞凋亡并抑制血管生成，其效力与RES相当，值得进一步评估用于癌症化学预防。
