Hubei Key Laboratory of Natural Products Research and Development, China Three Gorges University, Yichang 443002, China.
Molecules. 2019 Mar 20;24(6):1091. doi: 10.3390/molecules24061091.
The antitumor activity of CLE-10 (4-epi-isoinuviscolide), a sesquiterpene lactone compound, isolated from L. has rarely been reported. The aim of this study is to investigate the antitumor activity of CLE-10 and give a greater explanation of its underlying mechanisms.
The cytotoxicity of CLE-10 was evaluated using MTT assay. Autophagy was detected by the formation of mRFP-GFP-LC3 fluorescence puncta and observed using transmission electron microscopy, while flow cytometry was employed to detect apoptosis. The protein expressions were detected through Western blotting.
CLE-10 induced pro-death autophagy and apoptosis in MDA-MB-231 cells by increasing the protein expression of LC3-II, p-ULK1, Bax, and Bad, as well as downregulating p-PI3K, p-Akt, p-mTOR, p62, LC3-I, Bcl-2, and Bcl-xl. CLE-10 that was pretreated with 3-methyladenine (3-MA) or chloroquine (CQ) weakened the upregulation of the protein expression of p-ULK1, or the downregulation of p62, p-mTOR, and decreased the level of cytotoxicity against MDA-MB-231 cells. Meanwhile, rapamycin enhanced the effect of CLE-10 on the expression of autophagy-related protein and its cytotoxicity, with the IC value of CLE-10 decreasing from 4.07 µM to 2.38 µM.
CLE-10 induced pro-death autophagy and apoptosis in MDA-MB-231 cells by upregulating the protein expressions of LC3-II, p-ULK1, Bax, and Bad and downregulating p-PI3K, p-Akt, p-mTOR, p62, Bcl-2, and Bcl-xl.
从 L.中分离得到的倍半萜内酯化合物 CLE-10（4-表异土木香内酯）具有抗肿瘤活性，但鲜有报道。本研究旨在探讨 CLE-10 的抗肿瘤活性，并对其作用机制进行更深入的研究。
采用 MTT 法评价 CLE-10 的细胞毒性。采用 mRFP-GFP-LC3 荧光斑点形成法和透射电镜观察自噬，流式细胞术检测细胞凋亡。采用 Western blot 检测蛋白表达。
CLE-10 通过增加 LC3-II、p-ULK1、Bax 和 Bad 的蛋白表达，并下调 p-PI3K、p-Akt、p-mTOR、p62、LC3-I、Bcl-2 和 Bcl-xl 的表达，诱导 MDA-MB-231 细胞发生促死亡自噬和凋亡。用 3-甲基腺嘌呤（3-MA）或氯喹（CQ）预处理 CLE-10 可减弱 p-ULK1 蛋白表达的上调或 p62、p-mTOR 的下调，并降低对 MDA-MB-231 细胞的细胞毒性。同时，雷帕霉素增强了 CLE-10 对自噬相关蛋白表达及其细胞毒性的作用，使 CLE-10 的 IC 值从 4.07µM 降低至 2.38µM。
CLE-10 通过上调 LC3-II、p-ULK1、Bax 和 Bad 的蛋白表达，下调 p-PI3K、p-Akt、p-mTOR、p62、Bcl-2 和 Bcl-xl 的表达，诱导 MDA-MB-231 细胞发生促死亡自噬和凋亡。
