Research Center for Drug Discovery, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510006, People's Republic of China.
Research Center for Drug Discovery, School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou 510006, People's Republic of China.
Phytomedicine. 2022 Jul;101:154133. doi: 10.1016/j.phymed.2022.154133. Epub 2022 Apr 26.
Nasopharyngeal carcinoma (NPC) is highly prevalent in southern China. The remote metastasis of advanced NPC requires chemotherapeutic treatments to reduce the mortality. Our previous work revealed that saucerneol (SN) showed cytotoxicity against several nasopharyngeal carcinoma (NPC) cells. This work aims to investigate the effect of SN in NPC growth and exploring the mechanism of action.
Applying in vivo study, in vitro study and in silico study to indicate the mechanism of SN in inhibiting NPC growth.
Saucerneol (SN) toxicity was measured with MTT assay. NPC proliferation was measured with EdU and colony formation assays, cell cycle was detected with flow cytometry. NPC migration and invasion were measured with scratch assay and matrigel transwell method. Further, human NPC xenograft tumor models were established in nude mice to evaluate the therapeutic efficacy of SN in vivo. Toxicological analysis was performed on H&E staining and IHC. Quantitative real-time PCR and Western blot analyses were used to evaluate the expression levels of key molecules in PI3K/AKT/mTOR, MAPK, NF-κB, and HIF-1α signal pathways. Target predicting was conducted using computational method, and target identification was carried out by ATPase assay and TSA.
SN, a potent NPC inhibitor that was previously isolated from Saururus chinensis in our lab, is proven to inhibit the proliferation and metastasis of HONE1 cell lines and inhibit the growth of human NPC xenografts in nude mice. Moreover, we further articulate the molecular mechanism of action for SN and, reveal that SN promotes the expression of cell cycle-dependent kinase inhibitory protein p21 Waf1/Cip1 through targeting Grp94 and then inhibiting PI3K/AKT signaling pathway as well as up-regulating p53 to disrupt the progression of HONE1 cells.
SN significantly inhibits NPC cells proliferation and metastasis in vitro and in vivo via selectively inhibit Grp94 and then blocking PI3K/AKT/mTOR/HIF-1α signaling pathway. This study firstly provides a novel selective Grp94 inhibitor as a NPC candidate.
鼻咽癌（NPC）在中国南方高发。晚期 NPC 的远处转移需要化疗治疗以降低死亡率。我们之前的工作表明，獐牙菜苦醇（SN）对几种鼻咽癌（NPC）细胞具有细胞毒性。本研究旨在探讨 SN 对 NPC 生长的影响，并探索其作用机制。
应用体内研究、体外研究和计算机模拟研究来表明 SN 抑制 NPC 生长的机制。
采用 MTT 法测定獐牙菜苦醇（SN）的毒性。用 EdU 和集落形成实验测定 NPC 增殖，用流式细胞术检测细胞周期。用划痕实验和基质胶 Transwell 法测定 NPC 迁移和侵袭。进一步，在裸鼠中建立人 NPC 异种移植肿瘤模型，以评估 SN 的体内治疗效果。通过 H&E 染色和 IHC 进行毒理学分析。采用定量实时 PCR 和 Western blot 分析测定 PI3K/AKT/mTOR、MAPK、NF-κB 和 HIF-1α信号通路中关键分子的表达水平。通过计算方法进行靶标预测，并通过 ATP 酶测定和 TSA 进行靶标鉴定。
SN 是我们实验室之前从中华獐牙菜中分离得到的一种有效的 NPC 抑制剂，证明其能抑制 HONE1 细胞系的增殖和转移，并抑制裸鼠体内人 NPC 异种移植肿瘤的生长。此外，我们进一步阐明了 SN 的作用机制，并揭示 SN 通过靶向 Grp94 促进细胞周期依赖性激酶抑制剂 p21 Waf1/Cip1 的表达，从而抑制 PI3K/AKT 信号通路以及上调 p53 以破坏 HONE1 细胞的进展。
SN 通过选择性抑制 Grp94 并阻断 PI3K/AKT/mTOR/HIF-1α 信号通路，显著抑制 NPC 细胞在体外和体内的增殖和转移。本研究首次提供了一种新型的选择性 Grp94 抑制剂作为 NPC 候选药物。
