目的 研究β-榄香烯对人卵巢癌SKOV-3细胞凋亡的影响及机制的探索,从而为β-榄香烯在卵巢癌中的应用提供实验基础。方法 常规培养人卵巢癌SKOV-3细胞,传代,分组。应用Annexin V-FITC凋亡检测试剂盒及流式细胞仪以及TUNEL方法检测人卵巢癌SKOV-3细胞凋亡率,并采用Westen blot方法检测caspase 3、caspase 8、caspase 9、缺氧诱导因子-1α(HIF-1α)、Cyto-c及p-Akt蛋白表达。结果 与对照组相比,β-榄香烯处理组凋亡细胞数量和相关凋亡蛋白caspase-3、caspase-8、caspase-9及Cyto-c蛋白表达水平明显升高。与对照组相比,P-Akt及HIF-1α蛋白表达水平明显升高,而加入LY294002(PI3K/Akt/m TOR抑制剂),p-Akt及HIF-1α蛋白的表达水平明显降低。结论 β-榄香烯抑制宫颈癌细胞增殖和诱导细胞凋亡可能与上调凋亡蛋白及Cyto-c蛋白表达及活化PI-3K信号通路进而诱导HIF-1α高表达相关。
Objective To investigate the effects of β- -Elemene on apoptosis of human SKOV-3 cells and possible molecular mechanism. Methods The hmnan SKOV-3 cells were cultured and incubated with 40 μ g/ml or 80 μ g/ml β--Elemene for 24 hour. The cells in the control group were just incubated with medium. The expression of HIF- 1 α, p-Akt, Cyto-c, caspase3,caspase8 and caspase9 was detected by Western blot. Results β -elemene inhibited the viahility of human SKOV-3 cells in a dose-time dependent manner, induced the cell apoptosis. β-elemene up-regulaled tlle expression levels of Cyto-c, easpase3, caspase8, caspase9, p-Akt and HIF-1α, while co-treatment with PI3K/Akt/mTOR signaling pathway inhibitor LY294002 significantly down-regulated the expression level of HIF-1α. Conclusion β -elemene could increase the expression of HIF-1α through PI3K/Akt/mTOR signaling pathway, HIF-1α partially prevents human SKOV-3 cells from undergoing apoptosis.