dela Cruz Joseph Flores, Kim Yeon Soo, Lumbera Wenchie Marie Lara, Hwang Seong Gu
Department of Animal Life And Environmental Science, Molecular Biology And Biotechnology, Hankyong National University, Anseong-Si, Korea E-mail :
Asian Pac J Cancer Prev. 2015;16(15):6417-21. doi: 10.7314/apjcp.2015.16.15.6417.
Viscum album var (VAV) also known as mistletoe, has long been categorized as a traditional herbal medicine in Asia. In addition to its immunomodulating activities, mistletoe has also been used in the treatment of chronic hepatic disorders in China and Korea. There are numerous reports showing that VAV possesses anti-cancer effects, however influence on human hepatocarcinoma has never been elucidated. In the present study, hot water extracts of VAV was evaluated for its potential anti-cancer effect in vitro. SK-Hep1 cells were treated with VAV (50-400 ug/ml) for both 24 and 48 hours then cell viability was measured by cell counting kit-8 (CCK-8). Flow cytometry analysis was used to measure the proportion of SK-Hep1 in the different stages of cell cycle. RT-PCR and Western blot analysis were conducted to measure expression of cell cycle arrest related genes and proteins respectively. VAV dose dependently inhibited the proliferation of SK-Hep1 cells without any cytotoxicity with normal Chang liver cell (CCL-13). Flow cytometry analysis showed that VAV extract inhibited the cell cycle of SK-Hep1 cells via G1 phase arrest. RT-PCR and Western blot analysis both revealed that cyclin dependent kinase 2 (Cdk2) and cyclin D1 gene expression were significantly down regulated while p21 was upregulated dose dependently by VAV treatment. Combined down regulation of Cdk2, Cyclin D1 and up regulation of p21 can result in cell death. These results indicate that VAV showed evidence of anti-cancer activity through G1 phase cell cycle arrest in SK-Hep1 cells.
白果槲寄生变种（VAV）也被称为槲寄生，长期以来在亚洲被归类为传统草药。除了具有免疫调节活性外，槲寄生在中国和韩国还被用于治疗慢性肝脏疾病。有大量报道表明VAV具有抗癌作用，然而其对人类肝癌的影响尚未阐明。在本研究中，评估了VAV的热水提取物在体外的潜在抗癌作用。用VAV（50 - 400微克/毫升）处理SK - Hep1细胞24小时和48小时，然后通过细胞计数试剂盒 - 8（CCK - 8）测量细胞活力。采用流式细胞术分析来测量SK - Hep1细胞在细胞周期不同阶段的比例。分别进行RT - PCR和蛋白质印迹分析以测量细胞周期停滞相关基因和蛋白质的表达。VAV剂量依赖性地抑制SK - Hep1细胞的增殖，而对正常的张氏肝细胞（CCL - 13）没有任何细胞毒性。流式细胞术分析表明，VAV提取物通过G1期停滞抑制SK - Hep1细胞的细胞周期。RT - PCR和蛋白质印迹分析均显示，VAV处理可使细胞周期蛋白依赖性激酶2（Cdk2）和细胞周期蛋白D1基因表达显著下调，而p21则剂量依赖性地上调。Cdk2、细胞周期蛋白D1的联合下调和p21的上调可导致细胞死亡。这些结果表明，VAV通过使SK - Hep1细胞的细胞周期停滞在G1期而显示出抗癌活性的证据。
