The inhibitory effect of saponin A Ⅲ from Anemarrhena asphodeloides on Akirin2 gene expression and cell proliferation in liver cancer cells
Objective:To investigate the expression of Akirin2 in human liver cancer tissue and the potential mechanism of anemone saponin A Ⅲ(TA-Ⅲ)inhibiting the proliferation and promoting apoptosis of human liver cancer cells(HepG2).Methods:The RT-qPCR and immunohistochemistry was used to detect the expression of Akirin2 in 103 liver cancer tissues.in vitro experiment.The MTT method were used to detect the effects of Akirin2 overexpression group,Akirin2 inhibitor group,as well as different concentrations of TA-Ⅲ(0,5,10,20,40 mol/L)and paclitaxel group(80 g/ml)on the proliferation activity of HepG2 cells after intervention,plate cloning experiment.The flow cytometry was used to detect the effects of Akirin2 overexpression,Akirin2 inhibition,and TA-Ⅲ intervention on HepG2 cell cloning and apoptosis,respectively;Akirin2 and apoptotic protein were detected using the WB method.Treatment of HepG2 cells with TA-Ⅲ(40 mol/L)and simultaneous transfection of Akirin2 mimics(TA-Ⅲ + Akirin2 overexpression group)were performed to compare the effects of overexpression of Akirin2 on TA-Ⅲ intervention in cell proliferation and apoptosis.Results:Akirin2 was significantly overexpressed in the tissues of liver cancer patients(P<0.05),and the overexpression of Akirin2 was independently associated with poor prognosis in patients(P<0.05).The cell proliferation and cloning ability of the Akirin2 overexpression group were significantly higher than those of the control group(P<0.05),while the expression of apoptosis related proteins and cell apoptosis rate were lower than those of the control group(P<0.05).The cell proliferation and cloning ability of the Akirin2 inhibitor group were significantly lower than those of the control group(P<0.05),while the expression of apoptosis related proteins and cell apoptosis rate were higher than those of the control group(P<0.05).The proliferation activity of HepG2 cells in the TA-Ⅲ treatment group was significantly inhibited,and the inhibition of cell proliferation activity was proportional to the intervention concentration and time.The proliferation activity,cell cloning ability,and Akirin2 protein expression of HepG2 cells treated with TA-Ⅲ at different concentrations were significantly lower than those of the control group(P<0.05),and the cell apoptosis rate and apoptotic protein were significantly higher than those of the control group(P<0.05).After overexpression of Akirin2,the effects of TA-Ⅲ on the cell proliferation ability,apoptosis rate,and apoptosis related proteins of HepG2 cells were partially reversed.Conclusion:Akirin2 is significantly overexpressed in liver cancer,and TA-Ⅲ can inhibit cancer cell proliferation and promote apoptosis by inhibiting Akirin2 expression.
hepG2 cellsAnemarrhena saponin A Ⅲproliferationakirin2 apoptosis
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