謝逸憲李祥麟Yi-Hsien HsiehHsiang-Lin Lee楊子賢Zi-Xian Yang醫學研究所碩士班2026-10-292025-07-1020252025-06-23https://handle.ncl.edu.tw/11296/f8xre5https://scholars.csmu.edu.tw/handle/123456789/36733Chaetoglobosin A (ChA) 是一種從內生真菌 Chaetomium globosum Km1226 中分離出的代謝物，已顯示具有多種生物活性，包括抗腫瘤、抗真菌、植物毒性與免疫調節等作用。ChA可誘導多種癌細胞株的細胞毒性與凋亡。然而，ChA在人類肝細胞癌 (HCC) 中的抗癌機制尚不清楚。實驗結果顯示ChA顯著抑制肝細胞癌生長並誘導 G2/M 期細胞週期停滯，伴隨 p-Cdc2/Cyclin B1 的表現減少及γH2AX 表現增加。ChA同時誘發細胞凋亡、粒線體功能障礙與活性氧累積，並導致HO-1表現增加與Keap1表現下降。使用N-乙醯半胱氨酸 (NAC) 或siHO-1可減弱這些效應，顯示ROS–HO-1軸在ChA誘導的細胞凋亡中具有關鍵作用。RNA 定序結果進一步顯示，ChA抑制玻尿酸合成酶2 (HAS2)表現，此結果亦經由qPCR與西方墨點法確認。我們發現NAC處理可恢復細胞HAS2與RSK1的表現，顯示ChA可能透過 ROS–RSK1 路徑調控 HAS2 的表達。進一步實驗證實 ChA促進 RSK1 經由泛素–蛋白酶體途徑進行降解，進而導致下游 HAS2 表現下降。最後，ChA可與化療藥物 Regorafenib 和 Lenvatinib 協同增強對肝細胞癌的抑制效果。我們研究顯示ChA透過活化 ROS–HO-1並促進 RSK1 降解以抑制 HAS2 表達，誘導肝細胞癌的細胞週期停滯、凋亡與粒線體功能障礙，展現出抗癌作用。ChA有潛力成為肝細胞癌治療中的候選天然物。Chaetoglobosin A (ChA), a metabolite isolated from the endophytic fungus Chaetomium globosum Km1226, has demonstrated various biological activities including antitumor, antifungal, phytotoxic, and immunomodulatory effects. ChA induces cytotoxicity and apoptosis in various cancer cell lines. However, the anticancer mechanisms of ChA in human hepatocellular carcinoma (HCC) remain insufficiently characterized. Liver cancer cell lines were treated with varying concentrations of ChA. MTT assays. Colony formation assays, Annexin V staining, cell cycle analysis, reactive oxygen species (ROS) detection, mitochondrial potential assessments, siRNA knockdown experiments, RNA sequencing, and kinase array analysis to evaluate ChA's impact on HCC cellular functions. ChA significantly inhibited HCC cell growth and induced G2/M phase arrest, accompanied by downregulation of p-Cdc2/Cyclin B1 and upregulation of γ-H2AX. It also triggered apoptosis, mitochondrial dysfunction, and ROS accumulation, along with increased HO-1 and decreased Keap1 expression. N-acetylcysteine (NAC) treatment or HO-1 knockdown attenuated these effects, indicating a key role for the ROS–HO-1 axis in ChA-induced apoptosis. RNA sequencing further revealed downregulation of HAS2, a gene linked to hyaluronan synthesis and tumor progression, which was confirmed by qPCR and Western blot. Our results fund that NAC treatment restored the expression of both HAS2 and RSK1 in ChA-treated cells, suggesting that ChA may regulate HAS2 expression through an ROS–RSK1–dependent mechanism. Furthermore, proteasome inhibition by MG132 reversed the ChA-induced RSK1 downregulation, indicating that ChA promotes RSK1 degradation through the ubiquitin–proteasome pathway. Given that HAS2 expression was also restored under the same condition, our results suggest that ChA suppresses HAS2 expression at least in part by promoting RSK1 degradation. Notably, ChA synergized with Regorafenib and Lenvatinib to enhance anti-proliferative effects in HCC cells. Our findings demonstrate that ChA exerts potent anti-cancer effects in HCC cells by inducing cell cycle arrest, apoptosis, and mitochondrial dysfunction through activation of the ROS–HO-1 axis and inhibition of the RSK1–HAS2 signaling pathway. ChA may be as a potential therapeutic target for the treatment of HCC.肝細胞癌毛殼球菌素A活性氧細胞凋亡核糖體S6激酶玻尿酸合成酶Hepatocellular carcinoma cellsChaetoglobosin AROSCell apoptoticRSK1HAS2thesis