Saengboonmee Charupong, Thithuan Kanyarat, Mahalapbutr Panupong, Taebprakhon Cheerapinya, Aman Aamir, Rungrotmongkol Thanyada, Kamkaew Anyanee, Schevenels Florian Thierry, Chompupong Tanakiat, Wongkham Sopit, Lekphrom Ratsami
Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
Center for Translational Medicine, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.
Integr Cancer Ther. 2024 Jan-Dec;23:15347354241237519. doi: 10.1177/15347354241237519.
Hepatocellular carcinoma (HCC) is the most prevalent primary liver cancer. (Dunal) J.Sinclair (syn. ) has been used in Thai traditional medicine in various therapeutic indications. Phytochemical constituents of have been isolated and identified. However, their effects on liver cancer and the associated mechanisms have not been elucidated.
Dry flowers of were extracted using organic solvents, and chromatographic methods were used to purify the secondary metabolites. The chemical structures of the pure compounds were elucidated by analysis of spectroscopic data. Cytotoxicity against HCC cells was examined using SRB assay, and the effects on cell proliferation were determined using flow cytometry. The mechanisms underlying HCC inhibition were examined by molecular docking and verified by Western blot analysis.
Among 3 purified flavonoids, pinocembrin, pinostrobin, and chrysin, and 1 indole alkaloid (3-farnesylindole), only pinocembrin showed inhibitory effects on the proliferation of 2 HCC cell lines, HepG2 and Li-7, whereas chrysin showed specific toxicity to HepG2. Pinocembrin was then selected for further study. Flow cytometric analyses revealed that pinocembrin arrested the HCC cell cycle at the G1 phase with a minimal effect on cell death induction. Pinocembrin exerted the suppression of STAT3, as shown by the molecular docking on STAT3 with a better binding affinity than stattic, a known STAT3 inhibitor. Pinocembrin also suppressed STAT3 phosphorylation at both Tyr705 and Ser727. Cell cycle regulatory proteins under the modulation of STAT3, namely cyclin D1, cyclin E, CDK4, and CDK6, are substantially suppressed in their expression levels.
Pinocembrin extracted from exerted a significant growth inhibition on HCC cells suppressing STAT3 signaling pathways and its downstream-regulated genes.
肝细胞癌（HCC）是最常见的原发性肝癌。（杜纳尔）J. 辛克莱（同义词）已被用于泰国传统医学的各种治疗适应症。其植物化学成分已被分离和鉴定。然而，它们对肝癌的影响及其相关机制尚未阐明。
采用有机溶剂提取其干花，并采用色谱方法纯化次生代谢产物。通过光谱数据分析阐明纯化合物的化学结构。使用SRB法检测对肝癌细胞的细胞毒性，并使用流式细胞术测定对细胞增殖的影响。通过分子对接研究肝癌抑制的潜在机制，并通过蛋白质印迹分析进行验证。
在3种纯化的黄酮类化合物（松属素、松黄烷酮和白杨素）和1种吲哚生物碱（3 - 法呢基吲哚）中，只有松属素对两种肝癌细胞系HepG2和Li - 7的增殖有抑制作用，而白杨素对HepG2有特异性毒性。然后选择松属素进行进一步研究。流式细胞术分析表明，松属素使肝癌细胞周期停滞在G1期，对细胞死亡诱导的影响最小。松属素对STAT3有抑制作用，如分子对接显示其与已知的STAT3抑制剂stattic相比具有更好的结合亲和力。松属素还抑制了Tyr705和Ser727位点的STAT3磷酸化。在STAT3调控下的细胞周期调节蛋白，即细胞周期蛋白D1、细胞周期蛋白E、细胞周期蛋白依赖性激酶4（CDK4）和细胞周期蛋白依赖性激酶6（CDK6），其表达水平被显著抑制。
从其提取的松属素对肝癌细胞具有显著的生长抑制作用，抑制STAT3信号通路及其下游调控基因。
