冬凌草甲素诱导的铁死亡和细胞凋亡：双重抑制骨肉瘤细胞生长的策略。
Oridonin-induced ferroptosis and apoptosis: a dual approach to suppress the growth of osteosarcoma cells.
机构信息
Hunan University of Chinese Medicine, Changsha, China.
Henan Luoyang Orthopedic Hospital (Henan Provincial Orthopedic Hospital), Zhengzhou, China.
出版信息
BMC Cancer. 2024 Feb 12;24(1):198. doi: 10.1186/s12885-024-11951-1.
BACKGROUND
Osteosarcoma (OS) is one of the most common aggressive bone malignancy tumors in adolescents. With the application of new chemotherapy regimens, finding new and effective anti-OS drugs to coordinate program implementation is urgent for the patients of OS. Oridonin had been proved to mediate anti-tumor effect on OS cells, but its mechanism has not been fully elucidated.
METHODS
The effects of oridonin on the viability, clonal formation and migration of 143B and U2OS cells were detected by CCK-8, colony formation assays and wound-healing test. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was used to explore the mechanism of oridonin on OS. Western blot (WB), real-time quantitative PCR (qRT-PCR) were used to detect the expression levels of apoptosis and ferroptosis-relative proteins and genes. Annexin V-FITC apoptosis detection kit and flow cytometry examination were used to detect the level of apoptosis. Iron assay kit was used to evaluate the relative Fe content. The levels of mitochondrial membrane potential and lipid peroxidation production was determined by mitochondrial membrane potential detection kit and ROS assay kit.
RESULTS
Oridonin could effectively inhibit the survival, clonal formation and metastasis of OS cells. The KEGG results indicated that oridonin is associated with the malignant phenotypic signaling pathways of proliferation, migration, and drug resistance in OS. Oridonin was capable of inhibiting expressions of BAX, cl-caspase3, SLC7A11, GPX4 and FTH1 proteins and mRNA, while promoting the expressions of Bcl-2 and ACSL4 in 143B and U2OS cells. Additionally, we found that oridonin could promote the accumulation of reactive oxygen species (ROS) and Fe in OS cells, as well as reduce mitochondrial membrane potential, and these effects could be significantly reversed by the ferroptosis inhibitor ferrostatin-1 (Fer-1).
CONCLUSION
Oridonin can trigger apoptosis and ferroptosis collaboratively in OS cells, making it a promising and effective agent for OS therapy.
背景
骨肉瘤（OS）是青少年中最常见的侵袭性骨恶性肿瘤之一。随着新化疗方案的应用，迫切需要找到新的有效的抗 OS 药物来协调方案的实施。冬凌草甲素已被证明对 OS 细胞具有抗肿瘤作用，但作用机制尚未完全阐明。
方法
通过 CCK-8、集落形成实验和划痕愈合实验检测冬凌草甲素对 143B 和 U2OS 细胞活力、克隆形成和迁移的影响。京都基因与基因组百科全书（KEGG）富集分析用于探索冬凌草甲素对 OS 的作用机制。Western blot（WB）、实时定量 PCR（qRT-PCR）检测凋亡和铁死亡相关蛋白和基因的表达水平。Annexin V-FITC 凋亡检测试剂盒和流式细胞术检测凋亡水平。铁测定试剂盒用于评估相对 Fe 含量。线粒体膜电位检测试剂盒和 ROS 测定试剂盒分别用于测定线粒体膜电位和脂质过氧化产物水平。
结果
冬凌草甲素能有效抑制 OS 细胞的存活、克隆形成和转移。KEGG 结果表明，冬凌草甲素与 OS 中增殖、迁移和耐药等恶性表型信号通路相关。冬凌草甲素能抑制 143B 和 U2OS 细胞中 BAX、cl-caspase3、SLC7A11、GPX4 和 FTH1 蛋白和 mRNA 的表达，同时促进 Bcl-2 和 ACSL4 的表达。此外，我们发现冬凌草甲素能促进 OS 细胞中活性氧（ROS）和 Fe 的积累，降低线粒体膜电位，这些作用可被铁死亡抑制剂 Fer-1 显著逆转。
结论
冬凌草甲素能协同诱导 OS 细胞发生凋亡和铁死亡，是一种很有前途和有效的 OS 治疗药物。
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