Study of the effect of astragalin on proliferation of ovarian cancer cells by inhibiting the glycolytic pathway induced Via HIF-1α / 实用肿瘤学杂志

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 astra-galin 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 lym-phoma 2),Bax(Bcl-2-associated X protein),cleaved-caspase-3 and glycolysis related proteins such as Glut-1(Glucose trans-porter-1),Glut3,HK2(Hexokinase 2),PDK-1(Pyruvate dehydrogenase lipoamide kinase-1),PDK3 and HIF-1α(hypoxia-in-ducible 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 cul-ture 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.