熊果酸对喉鳞状细胞癌细胞周期阻滞的作用分析

Effect of ursolic acid on cell cycle arrest in laryngeal squamous cell carcinoma

目的探讨熊果酸对喉鳞状细胞癌细胞周期阻滞的作用。方法选取Hep-2喉鳞状细胞癌细胞接种于96孔板培养,依据随机数字表分为A组、B组、C组、D组,每组24孔,A组、B组、C组、D组分别给予0.0、12.5、25.0、50.0μmol/L熊果酸培养,采用Western blot(WB)法检测细胞周期相关调控蛋白周期蛋白(cyclin B1、cyclin D1)和周期蛋白依赖性激酶(CDK1、CDK2)表达情况,采用流式细胞术检测细胞周期,采用细胞增殖实验(MTT法)检测细胞增殖能力。结果在cyclin B1、cyclin D1、CDK1、CDK2的WB值及S期细胞占比中,D组明显低于C组,C组明显低于B组,B组明显低于A组,差异有统计学意义(P<0.05);在G0/G1期细胞占比中,D组明显高于C组,C组明显高于B组,B组明显高于A组,差异有统计学意义(P <0.05)。在培养1、2、3天的细胞增殖抑制率中,D组明显高于C组,C组明显高于B组,差异有统计学意义(P<0.05)。结论熊果酸可阻滞喉鳞状细胞癌细胞在G0/G1期细胞周期而抑制其细胞增殖,其机制作用可能够与下调cyclin B1、cyclin D1、CDK1、CDK2表达有关。

OBJECTIVE To discuss the effect of ursolic acid on cell cycle arrest in laryngeal squamous cell carcinoma cells.METHODS Hep-2 laryngeal squamous cell carcinoma cells were cultured on 96-well plate,they were divided into group A,group B,group C and group D according to the random number table,24 holes in each group,the group A,group B,group C and group D were cultured with 0.0,12.5,25.0,50.0μmol/L ursolic acid respectively.The Western blot(WB)was used to detect the expression of cyclin B1,cyclin D1 and cyclin dependent kinase(CDK1,CDK2),the cell cycle was detected by flow cytometry,the cell proliferation assay(MTT)was used to detect the cell proliferation ability.RESULTS In the cyclin B1,cyclin D1,CDK1,CDK2 WB value and S cell proportion,the group D was significantly lower than group C,group C was significantly lower than group B,group B was significantly lower than group A,the difference was statistically significant(P<0.05).In the G0/G1 cell proportion,the group D was significantly higher than group C,group C was significantly higher than group B,group B was significantly higher than group A,the difference was statistically significant(P<0.05).In the inhibition rate of cell proliferation after cultured for 1,2,3 days,the group D was significantly higher than group C,group C was significantly higher than group B,the difference was statistically significant(P<0.05).CONCLUSIONS Ursolic acid can arrest cell cycles in G0/G1 phase and inhibit cell proliferation of laryngeal squamous cell carcinoma cell.Its mechanism may be related to down-regulation of cyclin B1,cyclin D1,CDK1 and CDK2 expression.