The Action And Mechanism Of Ginsenoside Compound K On Cell Proliferation In Human Glioma Cell

Background and objectiveGlioma is the most common primary malignant tumour of the Central Nervous System,with a high morbidity and mortality rate.Despite treatment measures such as surgery,chemotherapy and radiotherapy were taken,the prognosis of patients remains poor.It is of great significance to overcome the limitations of existing therapies and find new drugs or treatments.In recent years,the anti-tumour effects of Chinese herbal monomers have received widespread attention.Ginsenoside Compound K(G-CK)is a rare ingredient extracted from the valuable medicinal herb ginseng.Studies have shown that G-CK has clear anti-tumour effects,but the role and mechanism of G-CK in glioma have not been fully elucidated.The aim of this study is to explore the role of G-CK in glioma and its possible molecular mechanisms.MethodsHuman glioma cells U251 and U87 were cultured in vitro.CCK-8 assay was used to explore the effect of series concentrations and times of G-CK on cell viability of U251 and U87 cells.Clone formation assay was connected to investigate the effect of G-CK on the proliferation of U251 cells.Immunofluorescence assay was performed to examine the effect of G-CK on Ki67 expression of U251 cells.In addition,the effect of G-CK on the migration of U251 cells was detected by cell damage healing assay.Western Blot was used to discover the effect of G-CK on the expression of related proteins,including Focal Adhesion Kinase(FAK)and Protein Kinase B(AKT/PKB).U251 and U87 were treated with Epidermal Growth Factor(EGF)and FAK inhibitor(PF-562271),and Western Blot further investigated the effects of single treatment with EGF and PF-562271,as well as co-incubation with G-CK+EGF and PF-562271+EGF on FAK/AKT signaling pathway in glioma cells.Finally,the effect of G-CK on EGF-induced cell proliferation was detected by CCK-8assay.Results1.G-CK inhibited the viability of glioma cells with dose-dependent and time-dependent effects.As the concentration or the duration of action increased,the absorbance of the G-CK group was decreased gradually(P<0.05).G-CK inhibited the clone formation of U251 cells.With the concentration of G-CK increased,the number of clones formed gradually decreased(P<0.05).G-CK inhibited Ki67 expression in U251 cells.As the concentration or the duration of action increased,the amount and intensity of Ki67 were decreased gradually(P<0.05).2.G-CK inhibited the migration of U251 cells,and the inhibition became more obvious with the increased of drug concentration and duration of action(P<0.05).3.Cells were treated with G-CK for the indicated times.FAK and AKT phosphorylation levels were measured by Western blot analysis.The results showed that with the prolonged effect of G-CK treatment on glioma cells,the phosphorylation expression of FAK and AKT decreased gradually(P<0.05).Next,Cells were further treated with G-CK and PF-562271 in the presence or absence of EGF,respectively,and the results showed that EGF upregulated basal levels of FAK and AKT phosphorylation,while G-CK significantly inhibited EGF-induced activation of the FAK/AKT phosphorylation pathway in glioma cells(P<0.05).4.Finally,Cells were treated with PF-562271 and EGF,and cell viability was measured by CCK-8 assay.The results showed that EGF promoted cell proliferation,and the cell viability enhanced by EGF could be significantly inhibited by G-CK(P<0.05);similarly,PF-562271 blocked FAK activity and inhibited the EGF-induced cell proliferation(P<0.05).Conclusion:1.G-CK can significantly inhibit the proliferation of glioma cells in a concentration-time-dependent manner.2.The mechanism of G-CK against glioma cells proliferation might be related to FAK/AKT pathway.3.G-CK is promising for the treatment of gliomas.