Mansonone F对人宫颈癌细胞增殖、凋亡、侵袭及迁移的影响

摘要

Objective To investigate the effects of Mansonone F on proliferation, apoptosis, invasion, and migration of human cervical cancer Hela cells and to explore the possible mechanism. Methods Hela cells in the logarithmic phase were selected and treated with 2.5, 5, and 10 μM Mansonone F for 24 h, respectively. The effects of Mansonone F on cell viability, apoptosis rate, invasion and migration abilities of Hela cells were detected by MTT assay, flow cytometry and Transwell assay, respectively. RT-PCR and Western blotting were used to detect the expression of focal adhesion kinase (FAK) mRNA, and the levels of FAK and phosphorylated FAK proteins. Results Mansonone F inhibited the proliferation of Hela cells at 24, 48, and 72 h, in a dose-dependent manner (P < 0.05). With the increase of drug concentration, the early and late apoptotic rates of Hela cells increased gradually, and the apoptotic rate at 10 μmol/L was higher than that at other concentrations (all P < 0.01). With the increase of drug concentration, the cell invasion and migration were markedly suppressed, and the number of penetrating cells was the lowest at 10 μmol/L (P < 0.01). Furthermore, Mansonone F significantly down-regulated the mRNA and protein expression of FAK, and the level of phosphorylated FAK protein after Hela cells were treated with 2.5, 5, and 10 μmol/L Mansonone F for 24 h. The lowest protein expression was observed at10 μmol/L (P < 0.01). Conclusion Mansonone F inhibits the proliferation, invasion, and migration, and induces the apoptosis of Hela cells by inhibiting the FAK expression, and down-regulating the level of phosphorylated FAK.%目的 观察Mansonone F对人宫颈癌Hela细胞体外增殖、凋亡、侵袭和迁移能力的影响, 并探讨其作用机制.方法 取对数生长期的Hela细胞, 分别加入2.5、5、10μmol/L Mansonone F培养.采用MTT法测算细胞增殖抑制率, 流式细胞术检测细胞凋亡率, Transwell小室实验观察细胞侵袭能力和细胞迁移能力, RT-PCR法检测细胞黏着斑激酶(FAK) mRNA表达, Western blotting法检测FAK、磷酸化黏着斑激酶(p-FAK) 蛋白表达.结果Mansonone F作用Hela细胞24、48、72 h后, 随着药物浓度的增加和作用时间的延长, 细胞增殖抑制率均升高(P均<0.05) .随着药物浓度的增加, 细胞早期凋亡率、晚期凋亡率均逐渐升高, 10μmol/L浓度下的细胞凋亡率高于其他各浓度(P均<0.01) .随着药物浓度的增加, 侵袭实验和迁移实验的穿膜细胞数目逐渐减少, 10μmol/L浓度下的穿膜细胞数最少(P <0.01);以2.5、5、10μmol/L Mansonone F作用Hela细胞24 h后, 随着药物浓度的升高, 细胞FAK mRNA及蛋白、p-FAK蛋白表达均降低, 10μmol/L作用下的细胞FAK mRNA及蛋白、p-FAK蛋白表达最低(P均<0.01) .结论 Mansonone F可显著抑制Hela细胞的增殖、诱导细胞凋亡, 抑制细胞侵袭和迁移, 其机制可能与抑制FAK表达、下调FAK的磷酸化水平有关.