Department of Hepatobiliary Surgery, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325035, China; Zhejiang-Germany Interdisciplinary Joint Laboratory of Hepatobiliary-Pancreatic Tumor and Bioengineering, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325035, China.
Department of Gastroenterology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325035, China.
Phytomedicine. 2024 Dec;135:156022. doi: 10.1016/j.phymed.2024.156022. Epub 2024 Sep 7.
Intrahepatic cholangiocarcinoma (ICC) is an aggressive and highly lethal cancer with an increasing incidence worldwide that lacks effective treatment regimens. Hypocrellin A (HA), a natural small compound isolated from S. bambusicola, has multiple biomedical activities, including antitumor activity.
We intended to investigate the therapeutic effects of HA on ICC and its potential mechanisms.
RBE and HuccT1 cell lines were utilized for in vitro experiments. CCK8 assay, colony formation analysis, RTCA, and immunofluorescence staining of ki67 were employed to evaluate the suppression effects of HA on proliferation. The inhibitory effects of HA on cell migration and invasion were evaluate through transwell and wound healing assays, and Hoechst 33,258 staining was performed to evaluate apoptosis. Additionally, we performed transcriptome sequencing and molecular docking for targeting identification, and immunoblotting and immunofluorescence of key molecules for validation. Two in vivo models, HuccT1 xenografts, and the primary ICC model (KRAS/P19/SB) established via hydrodynamic tail-vein injection were implemented. Multiplex immunohistochemistry (mIHC) was used to illustrate the multi-target inhibitory effects of HA.
The IC50 values of HA against RBE and HuccT1 cells were 4.612 μM and 10.01 μM for 24 h, as determined through the CCK8 assay. Our results confirmed that HA significantly repressed the proliferation, migration, invasion, and promoted the apoptosis of ICC cells at low concentrations. Moreover, HA exerted its anti-cancer effects through multi-target inhibition of the PI3K-AKT-mTOR, MAPK, and STAT3 signaling pathways. This inhibitory effect was rescued by Recilisib, an activator of the PI3K-AKT-mTOR pathway. Bioinformatics analysis of a multi-center RNA-Seq cohort (n = 90) demonstrated significant associations between these target pathways and the occurrence and poor prognosis of ICC. Animal studies suggested that HA strongly inhibited tumor growth in xenograft ICC models, and repressed the tumor number and size in the liver of primary ICC models by suppressing these three crucial pathways.
HA, a novel natural small molecule, demonstrated promising therapeutic efficacy against ICC through its multi-target inhibitory effects on the PI3K-AKT-mTOR, MAPK, and STAT3 signaling pathways. Moreover, it exhibited notable therapeutic benefits in a primary ICC model (KRAS/P19/SB), positioning it as a novel therapeutic agent for ICC.
肝内胆管癌（ICC）是一种侵袭性和高度致命的癌症，全球发病率不断上升，但缺乏有效的治疗方案。竹红菌甲素（HA）是一种从大青属中分离得到的天然小分子化合物，具有多种生物医学活性，包括抗肿瘤活性。
研究 HA 对 ICC 的治疗作用及其潜在机制。
采用 RBE 和 HuccT1 细胞系进行体外实验。通过 CCK8 检测、集落形成分析、RTCA 和 Ki67 免疫荧光染色评估 HA 对增殖的抑制作用。通过 Transwell 和划痕愈合实验评估 HA 对细胞迁移和侵袭的抑制作用，通过 Hoechst 33258 染色评估细胞凋亡。此外，我们进行了转录组测序和分子对接以确定靶点，并通过免疫印迹和关键分子的免疫荧光验证。建立 HuccT1 异种移植瘤和经尾静脉注射建立的原发性 ICC 模型（KRAS/P19/SB），进行了两项体内实验。多指标免疫组化（mIHC）用于说明 HA 的多靶点抑制作用。
CCK8 检测结果显示，HA 对 RBE 和 HuccT1 细胞的 IC50 值分别为 24 h 时的 4.612 μM 和 10.01 μM。结果证实，HA 以低浓度显著抑制 ICC 细胞的增殖、迁移和侵袭，并促进细胞凋亡。此外，HA 通过多靶点抑制 PI3K-AKT-mTOR、MAPK 和 STAT3 信号通路发挥抗癌作用。这种抑制作用可被 PI3K-AKT-mTOR 通路激活剂 Recilisib 挽救。对多中心 RNA-Seq 队列（n=90）的生物信息学分析表明，这些靶通路与 ICC 的发生和不良预后显著相关。动物研究表明，HA 强烈抑制异种移植 ICC 模型中的肿瘤生长，并通过抑制这三个关键通路抑制原发性 ICC 模型中的肿瘤数量和大小。
竹红菌甲素（HA）是一种新型天然小分子，通过多靶点抑制 PI3K-AKT-mTOR、MAPK 和 STAT3 信号通路，对 ICC 具有有前景的治疗效果。此外，它在原发性 ICC 模型（KRAS/P19/SB）中表现出显著的治疗益处，是一种新型的 ICC 治疗药物。
