Department of Pharmaceutical Engineering, School of Chemistry and Chemical Engineering, Southeast University, Nanjing, Jiangsu 211189, P.R. China.
Key Laboratory of Agro‑Products Processing Technology of Shandong Province, Institute of Agro‑Food Science and Technology, Shandong Academy of Agricultural Sciences, Jinan, Shandong 250100, P.R. China.
Mol Med Rep. 2019 May;19(5):3799-3806. doi: 10.3892/mmr.2019.10046. Epub 2019 Mar 15.
Ent‑3α‑formylabieta‑8(14),13(15)‑dien‑16,12β‑olide (EFLDO) is a compound extracted from Euphorbia lunulata Bge exhibiting anti‑proliferative activity in vitro. In the present study, EFLDO was identified to sensitize HepG2 cells to tumor necrosis factor (TNF) superfamily member 10 (TNFSF10)‑induced apoptosis. Liver cancer cells were resistant to TNFSF10; however, EFLDO increased TNFSF10‑induced cancer cell viability inhibition and cell apoptosis induction as assessed by MTT assay and Annexin V‑fluorescein isothiocyanate (FITC)/propidium iodide assay, respectively. The western blotting results suggested that treatment with EFLDO increased TNFSF10‑induced upregulation of the protein expression levels of pro‑apoptotic proteins, including BCL2 associated agonist of cell death, BCL2 associated X, apoptosis regulator, caspase‑3 (CASP3) and CASP8. Furthermore, treatment with EFLDO increased TNFSF10‑mediated downregulation of the protein expression level of the anti‑apoptotic protein BCL2 apoptosis regulator. Notably, the increase in the activity of CASP3 was consistent with the western blotting results. Treatment with EFLDO sensitized liver cancer cells to TNFSF10, and apoptosis was induced via the upregulation of TNF receptor superfamily member 10a (TNFRSF10A) and TNFRSF10B in a tumor protein p53 (p53)‑dependent manner, as detected by reverse transcription‑quantitative polymerase chain reaction and western blot analyses. In addition, p53 was identified to be necessary for EFLDO‑induced sensitivity to TNFSF10, as assessed by western blotting and Annexin V‑FITC assay. Collectively, the present results suggested a novel mechanism underlying EFLDO function in liver cancer. Treatment with EFLDO was able to increase the antitumor effect of TNFSF10 in liver cancer cells in a p53‑dependent manner.
依桐醇酸酯（EFLDO）是从大戟属植物月腺大戟中提取的一种化合物，具有体外抗增殖活性。在本研究中，EFLDO 被鉴定为可使 HepG2 细胞对肿瘤坏死因子（TNF）超家族成员 10（TNFSF10）诱导的细胞凋亡敏感。肝癌细胞对 TNFSF10 具有抗性；然而，EFLDO 增加了 TNFSF10 诱导的癌细胞活力抑制和细胞凋亡诱导，通过 MTT 测定和 Annexin V-荧光素异硫氰酸酯（FITC）/碘化丙啶测定分别进行评估。Western blot 结果表明，EFLDO 处理增加了 TNFSF10 诱导的促凋亡蛋白，包括 BCL2 相关细胞死亡激动剂、BCL2 相关 X、凋亡调节因子、半胱氨酸天冬氨酸蛋白酶 3（CASP3）和半胱氨酸天冬氨酸蛋白酶 8（CASP8）的蛋白表达水平上调。此外，EFLDO 处理增加了 TNFSF10 介导的抗凋亡蛋白 BCL2 凋亡调节因子蛋白表达水平下调。值得注意的是，CASP3 活性的增加与 Western blot 结果一致。EFLDO 使肝癌细胞对 TNFSF10 敏感，并通过肿瘤蛋白 p53（p53）依赖性方式诱导细胞凋亡，通过逆转录-定量聚合酶链反应和 Western blot 分析检测到 TNF 受体超家族成员 10a（TNFRSF10A）和 TNFRSF10B 的上调。此外，通过 Western blot 和 Annexin V-FITC 测定鉴定 p53 是 EFLDO 诱导对 TNFSF10 敏感性所必需的。综上所述，本研究结果提出了 EFLDO 在肝癌中的作用的新机制。EFLDO 处理能够以 p53 依赖的方式增加 TNFSF10 在肝癌细胞中的抗肿瘤作用。
