紫云英苷通过抑制HIF-1α诱导的糖酵解途径发挥抗卵巢肿细胞作用的研究

doi:10.11904/j.issn.1002-3070.2018.06.005

摘要/Abstract

摘要： 目的探讨紫云英苷在2 D及3 D细胞培养条件下对人卵巢癌OVCAR-8细胞的增殖作用及其可能机制。方法采用CCK-8试剂盒检测紫云英苷在2 D培养水平对OVCAR-8细胞增殖活力的作用;采用3 D细胞增殖活性检测试剂盒检测紫云英苷在3 D培养水平对OVCAR-8细胞增殖活力的作用;采用划痕实验检测紫云英苷对OVCAR-8细胞迁移能力的作用;采用凋亡试剂盒检测细胞凋亡水平;采用Western blot法在2 D及3 D培养条件下检测紫云英苷对OVCAR-8细胞细胞凋亡相关蛋白Bcl2(B-cell lymphoma 2)、Bax(Bcl-2-associated X protein)和cleaved-caspase-3以及糖酵解相关蛋白Glut(Glucose transporter)1、Glut3、HK2(Hexokinase 2)、PDK(Pyruvate dehydrogenase lipoamide kinase)1、PDK3以及HIF(Hypoxia-inducible factor)-1α的蛋白表达水平的影响;采用ELISA试剂盒检测HK2活性。结果 4~100 μmol/L紫云英苷在2 D培养条件下对OVCAR-8细胞增殖均有抑制作用, 且呈剂量-时间依赖效应关系(P＜0.05);紫云英苷可在3 D培养条件下明显抑制OVCAR-8细胞增殖(P＜0.05);同时, 紫云英苷可明显抑制OVCAR-8细胞迁移能力。此外, 紫云英苷处理后细胞凋亡水平明显增高, 且紫云英苷处理在2 D及3 D培养条件下均可抑制Bcl2、Glut1、Glut3、HK2、PDK1和PDK3蛋白的表达, 增加Bax及cleaved-caspase-3蛋白水平并抑制3 D培养诱导的HIF-1α的蛋白表达, 此外, 紫云英苷在2 D 培养条件下可抑制HK2活性, 均呈一定的剂量依赖效应关系(P＜0.05)。结论紫云英苷在2 D及3 D培养条件下对卵巢癌OVCAR-8细胞增殖均具有抑制作用。且可能通过抑制HIF-1α诱导的糖酵解通路以及激活线粒体凋亡通路。

关键词: 紫云英苷, 卵巢癌, 糖酵解, 凋亡, 3 D, 缺氧诱导因子

Abstract: Objective The Objective of this study was to explore the effect of astragalin on human ovarian cancer OVCAR-8 cells in 2 D and 3 D culture conditions and its possible mechanism. Methods CCK-8 assay was use to detect the effect of astragalin on the proliferation of OVCAR-8 cells in 2 D culture conditions.The 3 D cell proliferation activity assay kit was used to detect the effect of astragalin on OVCAR-8 cells in 3 D culture conditions.Cell apoptosis kit was used to detect the cell apoptosis rate after astragaline treatment.In addition, Western blot was used to detect the levels of apoptosis related proteins such as Bcl2(B-cell lymphoma 2), Bax(Bcl-2-associated X protein), cleaved-caspase-3 and glycolysis related proteins such as Glut-1(Glucose transporter-1), Glut3, HK2(Hexokinase 2), PDK-1(Pyruvate dehydrogenase lipoamide kinase-1), PDK3 and HIF-1α(hypoxia-inducible factor-1α)in OVCAR-8 cells after astragaline treatments in 2 D and 3 D culture conditions.HK2 activity was detected in OVCAR-8 cells by Elisa. Results Astragaline at doses of 4~100 μmol/L significantly inhibited the proliferation of OVCAR-8 cells in 2 D culture conditions, and showed a dose-and time-dependent manner(P＜0.05).Astragalin significantly inhibited the proliferation of OVCAR-8 cells in 3 D culture conditions(P＜0.05).Astragalin also significantly inhibited the migration ability of OVCAR-8 cells(P＜0.05).In addition, astragaline significantly increased apoptosis rate, decreased the levels of Bcl2, Glut1, Glut3, HK2, PDK1 and PDK3 proteins and increased the levels of Bax and cleaved-caspase-3 proteins in OVCAR-8 cells both in 2 D and 3 D culture conditions(P＜0.05).Astragaline significantly decreased the expression of HIF-1α in OVCAR-8 cells in 3 D culture conditions(P＜0.05).In addition, astragalin decreased HK2 activity in OVCAR-8 cells under 2 D culture conditions in a dose-dependent manner(P＜0.05). Conclusion Astragalin has an inhibitory effect on the proliferation of OVCAR-8 cells both in 2 D and 3 D culture conditions.Its mechanism may be related to inhibiting glycolytic and the mitochondrial apoptotic pathways induced by HIF-1α.

Key words: Astragaline, Ovarian cancer, Glycolysis, Apoptosis, 3 D, HIF-1α

中图分类号:

R737.31

宋玲, 付琼. 紫云英苷通过抑制HIF-1α诱导的糖酵解途径发挥抗卵巢肿细胞作用的研究[J]. 实用肿瘤学杂志, 2018, 32(6): 503-509.

SONG Ling, FU Qiong. Study of the effect of astragalin on proliferation of ovarian cancer cells by inhibiting the glycolytic pathway induced Via HIF-1α[J]. PRACTICAL ONCOLOGY JOURNAL, 2018, 32(6): 503-509.

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