Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China; State Key Laboratory of Research on Bioactivities and Clinical Applications of Medicinal Plants, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China.
Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China; State Key Laboratory of Research on Bioactivities and Clinical Applications of Medicinal Plants, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China; Department of Chemical Pathology, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China.
Phytomedicine. 2023 Aug;117:154900. doi: 10.1016/j.phymed.2023.154900. Epub 2023 May 30.
Patrinia villosa, a traditional medicinal herb commonly used for treating intestinal-related diseases, has been commonly prescribed by Chinese medicine practitioners as a key component herb to treat colon cancer, although its anti-tumor effect and mechanisms of action have not been fully elucidated.
HYPOTHESIS/PURPOSE: This study aimed to investigate the anti-tumor and anti-metastatic effects of Patrinia villosa aqueous extract (PVW), and its underlying mechanisms.
The chemical profile of PVW was analysed by high-performance liquid chromatography with photodiode-array detection (HPLC-DAD) method. Cell-based functional assays MTT, BrdU, scratch, and transwell were conducted to evaluate the effects of PVW on human colon cancer HCT116 and murine colon26-luc cells, assessing cytotoxicity, cell proliferation, motility, and migration, respectively. Western blotting was performed to assess the effect of PVW on the expression of key intracellular signaling proteins. In vivo studies were conducted using zebrafish embryos and tumor-bearing mice to evaluate the anti-tumor, anti-angiogenesis, and anti-metastatic effects of PVW in colon cancer.
Five chemical markers were identified and quantified in PVW. PVW exhibited significant cytotoxicity and anti-proliferative activity, as well as inhibitory effects on cell motility and migration in both HCT116 and colon 26-luc cancer cells via modulating protein expressions of TGF-β R1, smad2/3, snail, E-cadherin, FAK, RhoA, and cofilin. PVW (0.01-0.1 mg/ml) could significantly decrease the length of subintestinal vessels of zebrafish embryos through decreasing mRNA expressions of FLT1, FLT4, KDRL, VEGFaa, VEGFc, and Tie1. PVW (> 0.05 mg/ml) also significantly suppressed colon cancer cells migration in the zebrafish embryos. Furthermore, oral administration of PVW (1.6 g/kg) significantly inhibited tumor growth by decreasing the expressions of tumor activation marker Ki-67 and CD 31 in tumor tissues of HCT116 tumor-bearing mice. PVW could also significantly inhibit lung metastasis in colon 26-luc tumor-bearing mice by modulating their tumor microenvironment, including immune cells populations (T cells and MDSCs), levels of cytokines (IL-2, IL-12, and IFN-γ), as well as increasing the relative abundance of gut microbiota.
This study revealed for the first time the anti-tumor and anti-metastatic effects of PVW through regulation of TGF-β-smad2/3-E-cadherin, and FAK-cofilin pathways in colon cancer. These findings provide scientific evidence to support the clinical use of P. villosa in patients with colon cancer.
败酱草是一种传统的药用草本植物，常用于治疗肠道相关疾病，被中医医师广泛用作治疗结肠癌的关键草药。尽管其抗肿瘤作用及其作用机制尚未完全阐明。
假说/目的：本研究旨在探讨败酱草水提物（PVW）的抗肿瘤和抗转移作用及其潜在机制。
采用高效液相色谱-光电二极管阵列检测（HPLC-DAD）法分析 PVW 的化学成分。通过 MTT、BrdU、划痕和 Transwell 细胞功能测定法评估 PVW 对人结肠癌 HCT116 和鼠结肠癌 26-luc 细胞的影响，分别评估细胞毒性、细胞增殖、迁移和迁移能力。Western blot 用于评估 PVW 对关键细胞内信号蛋白表达的影响。通过斑马鱼胚胎和荷瘤小鼠进行体内研究，评估 PVW 在结肠癌中的抗肿瘤、抗血管生成和抗转移作用。
鉴定并定量了 PVW 中的 5 种化学标志物。PVW 通过调节 TGF-β R1、smad2/3、snail、E-cadherin、FAK、RhoA 和 cofilin 的蛋白表达，对 HCT116 和结肠 26-luc 癌细胞表现出显著的细胞毒性和抗增殖活性以及对细胞迁移和迁移的抑制作用。0.01-0.1mg/ml 的 PVW 可通过降低斑马鱼胚胎中 FLT1、FLT4、KDRL、VEGFaa、VEGFc 和 Tie1 的 mRNA 表达，显著减少亚肠血管的长度。0.05mg/ml 以上的 PVW 还可显著抑制斑马鱼胚胎中结肠癌细胞的迁移。此外，败酱草水提物（1.6g/kg）口服给药可通过降低肿瘤组织中肿瘤激活标志物 Ki-67 和 CD31 的表达，显著抑制 HCT116 荷瘤小鼠的肿瘤生长。PVW 还可通过调节免疫细胞群（T 细胞和 MDSC）、细胞因子（IL-2、IL-12 和 IFN-γ）水平以及增加肠道微生物群的相对丰度，显著抑制结肠 26-luc 荷瘤小鼠的肺转移。
本研究首次揭示了败酱草通过调节 TGF-β-smad2/3-E-cadherin 和 FAK-cofilin 通路在结肠癌中的抗肿瘤和抗转移作用。这些发现为败酱草在结肠癌患者中的临床应用提供了科学依据。
