Department of Orthopedics, Shanghai Bone Tumor Institution, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China.
Laboratory Animal Center, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200080, China.
J Ethnopharmacol. 2024 Dec 5;335:118709. doi: 10.1016/j.jep.2024.118709. Epub 2024 Aug 18.
Osteosarcoma (OS) is characterized by rapid growth and frequent pulmonary metastasis. Eurycoma longifolia Jack, a flowering plant primarily found in Southeast Asian countries, is commonly used in traditional herbal medicine. Its root extract is mainly used for against cancer, malaria, parasites and other conditions. The active compound in its root extract, eurycomanone (EUR), has been proven to inhibit lung and liver cancer proliferation.
Our research aimed to investigate the inhibitory effect and underlying molecular mechanism of EUR on OS growth and metastasis.
In vitro experiments: western blotting (WB) screened 41 compounds that inhibited GRP78 expression and evaluated the protein levels of GRP78, PARP, cleaved-PARP, MMP2, and MMP9. Cell proliferation was evaluated using CCK-8, EdU, colony formation assay, and cell apoptosis was assessed by flow cytometry. Transwell, wound healing, and tube formation assays were performed to determine the effect of EUR on tumor invasion, migration, and angiogenesis, respectively. Quantitative real-time polymerase chain (qRT-PCR) and dual-luciferase activity assays detected GRP78 mRNA stability and transcription levels post-EUR and thapsigargin treatment. RNA-Seq identified signaling pathways inhibited by EUR. In vivo experiments: effects of EUR in mice were evaluated by H&E staining to detect lung metastasis and potential toxic effects in tissues. Immunohistochemical (IHC) staining detected the expression of Ki-67, CD31, and cleaved caspase-3 in tumors.
GRP78 is highly expressed in OS and correlated with poor prognosis. In vitro, eurycomanone (EUR) significantly downregulated GRP78 expression, inhibited cell proliferation, migration, invasion, tube formation, and induced apoptosis. Moreover, it enhanced trichostatin A (TSA) sensitivity and exhibited inhibitory effects on other cancer types. Mechanistically, EUR decreased GRP78 mRNA stability and transcription. In vivo, EUR inhibited proliferation and invasion in tibial and PDX models.
Our study demonstrated that EUR inhibits the growth and metastasis of OS by reducing GRP78 mRNA stability and inhibiting its transcription, which offers a novel approach for clinical treatment of OS.
骨肉瘤（OS）的特征是生长迅速，经常发生肺转移。长叶补肾草，一种主要生长在东南亚国家的开花植物，常用于传统草药。其根提取物主要用于抗癌、疟疾、寄生虫等疾病。其根提取物中的活性化合物，eur ycoma none（EUR），已被证明能抑制肺癌和肝癌的增殖。
我们的研究旨在探讨 EUR 对 OS 生长和转移的抑制作用及其潜在的分子机制。
体外实验：western blot（WB）筛选出 41 种抑制 GRP78 表达的化合物，并评估了 GRP78、PARP、cleaved-PARP、MMP2 和 MMP9 的蛋白水平。用 CCK-8、EdU、集落形成实验评估细胞增殖，用流式细胞术评估细胞凋亡。Transwell、划痕愈合和管形成实验分别用于测定 EUR 对肿瘤侵袭、迁移和血管生成的影响。定量实时聚合酶链反应（qRT-PCR）和双荧光素酶活性测定检测 EUR 和 thapsigargin 处理后 GRP78 mRNA 的稳定性和转录水平。RNA-Seq 鉴定了 EUR 抑制的信号通路。体内实验：通过 H&E 染色检测肺转移和组织中潜在的毒性作用来评估 EUR 在小鼠中的作用。免疫组织化学（IHC）染色检测肿瘤中 Ki-67、CD31 和 cleaved caspase-3 的表达。
GRP78 在 OS 中高表达，与预后不良相关。体外，eur ycoma none（EUR）显著下调 GRP78 表达，抑制细胞增殖、迁移、侵袭、管形成和诱导细胞凋亡。此外，它增强了 trichostatin A（TSA）的敏感性，并对其他癌症类型具有抑制作用。在机制上，EUR 降低了 GRP78 mRNA 的稳定性和转录。在体内，EUR 抑制了胫骨和 PDX 模型中的增殖和侵袭。
我们的研究表明，EUR 通过降低 GRP78 mRNA 的稳定性和抑制其转录来抑制 OS 的生长和转移，为 OS 的临床治疗提供了一种新的方法。
