摘要
目的 探讨小豆蔻明(CAR)对宫颈癌HeLa细胞恶性进展的影响及其调控机制.方法 体外培养人宫颈癌HeLa细胞并随机分为对照组(Control组)、CAR低浓度组(CAR L组,10 μmol/L CAR)、CAR高浓度组(CAR H组,40 μmol/L CAR)和 CAR 高浓度+Akt 激活剂组(CAR H+SC79 组,40 μmol/L CAR+10 μmol/L SC79),Hoechst33258染色法观察各组细胞形态;CCK-8法检测细胞活力;细胞划痕实验检测细胞迁移率;Transwell实验检测细胞侵袭数;流式细胞术检测细胞凋亡;Western blot技术检测p-Akt、Akt、p-MDM2、MDM2、p-p53、p53蛋白表达水平.结果 与Control组比较,CARL、CAR H组细胞核皱缩、裂解,细胞增殖、迁移和侵袭及p-Akt/Akt、p-MDM2/MDM2水平下降,细胞凋亡率和p-p53/p53水平升高(P＜0.05);与CARH组比较,CAR H+SC79组细胞生长状态好转,细胞增殖、迁移和侵袭及p-Akt/Akt、p-MDM2/MDM2水平升高,细胞凋亡率和p-p53/p53水平下降(P＜0.05).结论 CAR通过调节Akt/MDM2/p53信号通路抑制宫颈癌HeLa细胞增殖、迁移与侵袭,诱导细胞凋亡.
Abstract
Objective To explore the effect and regulatory mechanism of cardamonin(CAR)on the malignant progression of HeLa cells of cervical cancer.Methods Human cervical cancer HeLa cells were cultured in vitro and randomly grouped into a Control group,a low concentration CAR group(CAR L group,10 μmol/L CAR),a high concentration CAR group(CAR H group,40 μmol/L CAR),and a high concentration CAR+Akt activator group(CAR H+SC79 group,40 μmol/L CAR+10 μmol/L SC79),Hoechst33258 staining method was applied to observe the cell morphology of each group.CCK-8 method was applied to detect cell viability.Cell scratch test was applied to detect cell migration rate.Transwell experiment was applied to detect the number of cell invasions.Flow cytometry was applied to detect cell apoptosis.Western blot technol-ogy was applied to detect the expression levels of p-Akt,Akt,p-MDM2,MDM2,p-p53,and p53 proteins.Results Compared with the Control group,the nuclei of the CAR L and CAR H groups were shrunk and lysed,the cell proliferation,migration,and invasion,and the levels of p-Akt/Akt,p-MDM2/MDM2 were decreased,the rate of cell apoptosis and the level of p-p53/p53 were increased(P＜0.05).Compared with the CAR H group,the cell growth status of the CAR H+SC79 group was improved,the cell proliferation,migration,and invasion,and the levels of p-Akt/Akt,p-MDM2/MDM2 were increased,the rate of cell apoptosis and the level of p-p53/p53 were decreased(P＜0.05).Conclusion CAR inhibitde the proliferation,migra-tion,and invasion of HeLa cells of cervical cancer,and induced cell apoptosis by regulating the Akt/MDM2/p53 signaling pathway.
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