粗毛豚草素对人肺癌细胞A-549的抑制作用研究  

Inhibiting Effects of Hispidulin on Lung Carcinoma A-549 Cell Line

作　　者：张成军[1] 邓雁如[2] 

机构地区：[1]天津中医药大学第二附属医院呼吸科,300150 [2]天津中医药大学

出　　处：《天津医药》2013年第5期430-433,共4页Tianjin Medical Journal

摘　　要：目的探讨粗毛豚草素体外对人肺癌细胞A-549增殖的抑制作用及其作用机制。方法粗毛豚草素作用人肺癌细胞A-549后,应用MTT法检测细胞增殖;流式细胞术检测细胞周期;Western blot检测周期蛋白cyclin D1,细胞周期蛋白依赖性激酶(CDK)4,细胞周期蛋白依赖性激酶抑制蛋白p21和p27的表达;免疫细胞化学技术检测增殖细胞核抗原(PCNA)的阳性表达。结果 A-549细胞经不同浓度(15、30、60、120μmol/L)粗毛豚草素作用72h后,细胞存活率分别为(57.30±3.92)%、(43.91±4.32)%、(17.82±2.73)%和(13.20±2.46)%,均低于对照组(96.30±2.08)%,差异有统计学意义(P﹤0.01)。粗毛豚草素(15、30μmol/L)作用A-549细胞24h,可使细胞发生G1/S期阻滞。3种浓度(15、30、60μmol/L)的粗毛豚草素作用24h,可抑制A-549细胞周期蛋白cyclinD1、CDK4的表达,使p21蛋白表达增高,但对p27蛋白表达无明显影响。粗毛豚草素(15、30、60μmol/L)可显著抑制PCNA表达,其表达强度分别为(35.7±2.3)%、(26.6±2.1)%和(14.1±2.2)%,均低于对照组(45.2±3.3)%(P﹤0.01)。结论粗毛豚草素可抑制肺癌细胞A-549增殖,其机制可能与改变细胞周期分布,下调cyclinD1、CDK4表达,上调p21的表达等有关。Objective To investigate the effects of hispidulin on the proliferation of lung carcinoma A-549 cell line, and the mechanism thereof. Methods After human lung carcinoma cell line A-549 was treated with different concentrations of hispidulin, the cellular proliferation was detected by MTT assay. The flow cytometry was used to determine the cell cycle in A-549 cells. Western blot method was employed to detect the expressions of cyelin D1, eyelin-dependent kinase （CDK）4, cy- clin-dependent kinase inhibitor p21 and p27 in A-549 cells. Immunocytochemistry was used to detect the expression of pro- liferation cell nuclear antigen （PCNA） in A-549 ceils. Results The proliferation of lung carcinoma cells was significantly inhibited by hispidulin. The cell viability values were （57.30±3.92）%, （43.91±4.32）%, （17.82±2.73）% and （13.20±2.46）% re- spectively after treatment with the concentrations of hispidulin 15, 30, 60 and 120 μmol/L for 72 h, which were significantly lower than those （96.30±2.08）% in control group （P〈 0.01）. After treatment with hispidulin （15 and 30 μmol/L ） for 24 h, GJS phase cell cycle arrest was induced in A-549 cells. The expression levels of cyclin D1 and CDK4 proteins were inhibited in A-549 cells after treatment with hispidulin （15, 30 and 60 μmol/L ） for 24 h, but the expression of p21 protein was increased and the expression of p27 protein was not changed. The expression of PCNA was significantly inhibited by hispidulin （15, 30 and 60 μmol/L ） with the expressive rates （35.7±2.3）%, （26.6±2.1）% and （14.1±2.2）%, which were significantly lower than those of control group （P 〈 0.01）. Conclusion Hispidulin can inhibit the proliferation of A-549 cells through changing cell cycle, which may be related to the up-regulated level of p21 and down-regulated levels of eyclin D 1 and C DK4.

关 键 词：肺肿瘤 细胞系 肿瘤 细胞增殖 细胞周期 细胞周期蛋白D1 细胞周期蛋白依赖激酶4 周期素依赖 激酶抑制剂p21 周期素依赖激酶抑制剂P27 增殖细胞核抗原 粗毛豚草素 

分 类 号：R734.2[医药卫生—肿瘤]