Effects of Gypenoside on the malignant biological behaviors of oral squamous cell carcinoma cells by regulating the PKA/CREB signaling pathway
JIN Jie
LU Ping
WEI Zhen
Abstract：Objective To investigate the effects of Gypenoside(GYP)on the malignant biological behaviors of oral squamous cell carcinoma(OSCC)cells by regulating the protein kinase A(PKA)/cAMP-response element binding protein(CREB)signaling pathway.Methods Human oral squamous cell carcinoma cells(HSC3)were treated with GYP at a concentration of 2.5-160mg/L.The cell counting kit-8(CCK-8)assay was applied to detect cell activity and screen for the optimal drug concentration.HSC3 cells were induced with blank control,low-dose,medium-dose and high-dose GYP,and high-dose GYP+Sp-cAMP(PKA activator).Cell proliferation,apoptosis,migration and invasion were detected by colony formation assay,flow cytometry,wound healing assay and Transwell assay,respectively.Western blot was applied to detect protein levels of cell proliferation antigen markers(Ki67,cyclin D1,and Caspase-3),B-cell lymphoma associated X-protein(Bax),matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9),protein kinase A(PKA),phosphorylated protein kinase A(p-PKA),and cAMP-response element binding protein(CREB)phosphorylated-cAMP-response element binding protein(p-CREB).An in vivo OSCC model was created in nude mice bearing OSCC xenografts,thus detecting the effect of GYP on the in vivo growth of OSCC.Results GYP concentrations of 20.0mg/L,40.0mg/L,and 80.0mg/L were selected as the low,medium and high doses for subsequent experiments,respectively.Low-dose,medium-dose and high-dose GYP treatment significantly decreased the number of colonies,wound healing rate,invasive cell number,and protein levels of Cyclin D1,Ki67,MMP-2,MMP-9,p-PKA/PKA,and p-CREB/CREB,but significantly increased the apoptotic rate,and protein levels of Caspase-3 and Bax than the blank control(P＜0.05).High-dose GYP+Sp-cAMP treatment significantly increased the number of colonies,wound healing rate,invasive cell number,and protein levels of Cyclin D1,Ki67,MMP-2,MMP-9,p-PKA/PKA,and p-CREB/CREB,but significantly decreased the apoptotic rate,and protein levels of Caspase-3 and Bax than those induced with high-dose GYP(P＜0.05).In vivo data showed that GYP-induced nude mice bearing OSCC xenografts had significantly lower tumor volume and weight,p-PKA/PKA,and p-CREB/CREB than the negative controls(P＜0.05).Conclusion GYP can inhibit the malignant biological behaviors of OSCC cells by inhibiting the PKA/CREB signaling pathway.
Keywords：oral squamous cell carcinomaGypenosideprotein kinase A(PKA)/cAMP-response element binding protein(CREB)malignant biological behavior
Publication Date：2025-03-26
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 375-380 )