葡萄籽多酚通过糖酵解和细胞周期阻滞抑制口腔鳞癌细胞的增殖和迁移

Grape Seed Polyphenols Inhibit Proliferation and Migration of Oral Squamous Cell Carcinoma Cells Via Reduced Glycolysis and Cell Cycle Arrest

目的探究葡萄籽多酚(GSP)对人口腔鳞癌细胞(OSCC)增殖、迁移、周期及糖酵解的影响及机制。方法使用3、6、12、24、48μg·mL^(-1)的GSP处理口腔鳞癌细胞株(HN6及Cal27)。采用CCK8法检测细胞增殖,划痕实验检测细胞迁移,流式细胞术检测细胞周期,葡萄糖氧化酶法检测细胞上清中葡萄糖含量,比色法检测丙酮酸和乳酸含量,RT-qPCR法检测细胞增殖、迁移及周期相关基因表达水平。结果GSP在6~48μg·mL^(-1)范围内对HN6及Cal27细胞株的增殖和迁移均具有显著的抑制作用,且存在剂量依赖性(P<0.05或P<0.01)。GSP处理的HN6细胞被阻滞于G1期,而Cal27细胞则被阻滞于G2期。与对照组相比,GSP处理组能够显著降低OSCC细胞的葡萄糖摄取、丙酮酸和乳酸的产生(P<0.01);GSP处理组肿瘤细胞增殖(MKi67、MYC、PCNA)、迁移(CDH2、Twist、Slug)和细胞周期(CyclinD1、CDK4、CDK6、CyclinB2、CDK1)等相关基因的表达水平显著降低(P<0.01)。结论GSP能抑制OSCC细胞的增殖、迁移,其机制可能与糖酵解降低和细胞周期G1期或G2期阻滞相关。

Objective To elucidate the effects of grape seed polyphenols(GSP)on the proliferation,migration,cell cycle,and glycolysis of human oral squamous cell carcinoma(OSCC)cells and its underlying mechanisms.Methods Oral squamous cell carcinoma cell lines(HN6 and Cal27)were treated with GSP at concentrations of 3,6,12,24,and 48μg·mL^(-1);cell proliferation was evaluated using the CCK8 assay,while cell migration was assessed via the scratch assay;flow cytometry was employed to analyze cell cycle progression;glucose concentrations in the cell supernatant were quantified using the glucose oxidase method;pyruvate and lactate concentrations were determined through colorimetric assays;the expression levels of genes associated with cell proliferation,migration,and cell cycle were measured using RT-qPCR.Results GSP demonstrated a significant inhibitory effect on the proliferation and migration of HN6 and Cal27 cell lines within the concentration range of 6 to 48μg·mL^(-1),exhibiting a dose-dependent relationship(P<0.05 or P<0.01).Treatment with GSP induced cell cycle arrest in the G1 phase in HN6 cells and in the G2 phase in Cal27 cells.Compared to the control group,the GSP-treated group exhibited a significant reduction in glucose uptake,pyruvate production,and lactate production in OSCC cells(P<0.01).Furthermore,GSP treatment resulted in a significant downregulation of the expression levels of genes associated with tumor cell proliferation(MKi67,MYC,PCNA),migration(CDH2,Twist,Slug),and cell cycle(CyclinD1,CDK4,CDK6,CyclinB2,CDK1)compared to the control group(P<0.01).Conclusions GSP can inhibit the proliferation and migration of OSCC cells,potentially through the mechanisms involving reduced glycolysis and cell cycle arrest in the G1 or G2 phase.