绞股蓝皂苷调节PKA/CREB信号通路对口腔鳞癌细胞恶性生物学行为的影响  

Effects of Gypenoside on the malignant biological behaviors of oral squamous cell carcinoma cells by regulating the PKA/CREB signaling pathway

作　　者：金婕 卢平 魏震 徐树森 JIN Jie;LU Ping;WEI Zhen(Department of Children's Stomatology,Changsha Stomatological Hospital,Hunan,Changsha 410000,China)

机构地区：[1]湖南省长沙市口腔医院儿童口腔科,410000

出　　处：《河北医药》2025年第3期375-380,共6页Hebei Medical Journal

基　　金：湖南省卫生健康委科研计划课题(编号:D202308016829)。

摘　　要：目的探究绞股蓝皂苷(GYP)调节蛋白激酶A(PKA)/环磷腺苷效应元件结合蛋白(CREB)信号通路对口腔鳞状细胞癌(OSCC)细胞恶性生物学行为的影响。方法用浓度为2.5~160 mg/L的GYP处理人口腔鳞癌细胞(HSC3),CCK-8法用以检测细胞活性,筛选GYP浓度;将HSC3细胞分为对照组(Control组),绞股蓝皂苷低(GYP-L组)、中(GYP-M组)、高浓度组(GYP-H组)、高浓度+PKA激活剂组(GYP-H+Sp-cAMP组);采用平板克隆法检测细胞增殖;用流式细胞仪检测细胞凋亡;用划痕实验检测细胞迁移;Transwell实验检测细胞侵袭;Western blot检测细胞核增殖抗原标记物(Ki67)、细胞周期蛋白D1(Cyclin D1)、半胱氨酸蛋白酶-3(Caspase-3)、B细胞淋巴瘤-2相关X蛋白(Bax)、基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、蛋白激酶A(PKA)、磷酸化蛋白激酶A(p-PKA)、环磷腺苷效应元件结合蛋白(CREB)、磷酸化环磷腺苷效应元件结合蛋白(p-CREB)蛋白表达;裸鼠移植瘤检测GYP对OSCC移植瘤生长的影响。结果选择GYP浓度为20.0 mg/L、40.0 mg/L、80.0 mg/L进行后续实验。GYP-L、GYP-M、GYP-H组集落形成数、划痕愈合率、侵袭数量、Cyclin D1、Ki67、MMP-2、MMP-9、p-PKA/PKA、p-CREB/CREB表达水平比Control组低,细胞凋亡率、Caspase-3、Bax表达水平比Control组高(P<0.05);GYP-H+Sp-cAMP组集落形成数、划痕愈合率、侵袭数量、Cyclin D1、Ki67、MMP-2、MMP-9、p-PKA/PKA、p-CREB/CREB表达水平比GYP-H组高,细胞凋亡率、Caspase-3、Bax表达水平比GYP-H组低(P<0.05)。体内实验显示,GYP组小鼠肿瘤体积、质量减小,p-PKA/PKA、p-CREB/CREB水平下降(P<0.05)。结论GYP可通过抑制PKA/CREB信号通路抑制OSCC细胞恶性生物学行为。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

关 键 词：口腔鳞状细胞癌 绞股蓝皂苷 蛋白激酶A/环磷腺苷效应元件结合蛋白 恶性生物学行为 

分 类 号：R739.8[医药卫生—肿瘤]