Department of Pharmaceutical Engineering, School of Chemistry and Chemical Engineering, Southeast University, Nanjing, 211189, PR China.
Department of Pharmaceutical Engineering, School of Chemistry and Chemical Engineering, Southeast University, Nanjing, 211189, PR China; Jiangsu Province Hi-Tech Key Laboratory for Biomedical Research, Southeast University, PR China.
Biomed Pharmacother. 2018 Apr;100:407-416. doi: 10.1016/j.biopha.2018.02.004. Epub 2018 Feb 16.
To study the apoptosis induced by EFLDO (ent-3α-formylabieta-8(14), 13(15)-dien-16,12β-olide), extracted from the Euphorbia lunulata Bge, in the HepG2 cell line and to study the antitumor activity of this compound in vivo, Cell viability and migration were evaluated with CCK-8 (2-(2-methoxy-4-nitrophenyl)-3- (4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt) and wound healing assays, respectively. In addition, the cell cycle was examined using flow cytometry after propidium iodide (PI) staining. Apoptosis was analyzed by using the Annexin V/PI staining assay. Pro-caspase activation and apoptosis protein expression were evaluated by western blotting. A HepG2 xenograft model in nude mice was also established to study the antitumor activity of EFLDO in vivo. Immunohistochemical analysis was used to detect the expression of Ki67 in the tumors in situ. EFLDO could induce dose- and time-dependent apoptosis in HepG2 human hepatic cancer cells. Activation of caspases 3, 8, and 9 played an important role in EFLDO-induced apoptosis in vitro. Decreased levels of Bcl-2 and Survivin and increased level of BAX were also involved in this process. Furthermore, EFLDO could inhibit HepG2 tumor growth in nude mice, and the proliferation characteristics, reflected by the Ki67 index, were suppressed significantly. The results indicated that EFLDO could induce apoptosis in hepatic cancer cells by caspase activation in vitro and suppress tumor growth in vivo.
为了研究从Euphorbia lunulata Bge 中提取的 EFLDO（ent-3α-formylabieta-8(14),13(15)-dien-16,12β-olide）诱导 HepG2 细胞系细胞凋亡的作用，并研究该化合物在体内的抗肿瘤活性，用 CCK-8（2-(2-甲氧基-4-硝基苯基)-3-(4-硝基苯基)-5-(2,4-二磺基苯基)-2H-四唑，单钠盐）和划痕愈合实验分别评估细胞活力和迁移，另外，用碘化丙啶（PI）染色后用流式细胞术检查细胞周期。用 Annexin V/PI 染色实验分析细胞凋亡。用 Western blot 检测原半胱天冬酶激活和凋亡蛋白表达。还建立了 HepG2 荷瘤裸鼠模型，以研究 EFLDO 在体内的抗肿瘤活性。免疫组织化学分析用于检测原位肿瘤中 Ki67 的表达。EFLDO 可诱导 HepG2 人肝癌细胞呈剂量和时间依赖性凋亡。体外实验中，caspase-3、8 和 9 的激活在 EFLDO 诱导的细胞凋亡中起重要作用。Bcl-2 和 Survivin 水平降低，BAX 水平升高也参与了这一过程。此外，EFLDO 可抑制裸鼠 HepG2 肿瘤生长，Ki67 指数显著抑制肿瘤的增殖特征。结果表明，EFLDO 可通过体外 caspase 激活诱导肝癌细胞凋亡，并抑制体内肿瘤生长。
