天津医药

• 论著 • 上一篇    下一篇

粗毛豚草素对人肺癌细胞A-549的抑制作用研究

张成军1,邓雁如2   

  1. 1. 天津中医药大学第二附属医院
    2. 天津中医药大学
  • 收稿日期:2012-08-06 修回日期:2012-11-23 出版日期:2013-05-15 发布日期:2013-05-15
  • 通讯作者: 邓雁如

Effects of Hispidulin on Inhibiting Lung Carcinoma A549 Cell Line

ZHANG Cheng jun1,DENG YAN RU2   

  1. 1. The Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine, Tianjin 300150, China
    2. TianjinUniversity of Traditional Chinese Medicine
  • Received:2012-08-06 Revised:2012-11-23 Published:2013-05-15 Online:2013-05-15
  • Contact: DENG YAN RU

PDF (PC)

2515

摘要： 【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。

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

Abstract: Abstract   Objective  To investigate the effects and the mechanism of hispidulin on the proliferation of lung carcinoma A549 cell line. Method   After human lung carcinoma cell line A549 was treated with different concentrations of hispidulin, the cellular proliferation was detected by MTT assay. The flow cytometry was used to determine the change of cell cycle in A-549 cells. Western blot was employed to detect the expressions of cyclin D1, CDK4, p21 and p27 in A-549 cells. Immunocytochemistry was used to detect the expression of proliferation cell nuclear antigen in A-549 cells. Result  The proliferation of lung carcinoma cells was inhibited significantly by hispidulin. The cell viability values were (57.30±3.92)%, (43.91±4.32)%, (17.82±2.73)% and (13.20±2.46)% respectively after treatment with the concentrations of hispidulin 15、30、60、120μmol?L-1 for 72h, which were significantly lower than those (96.30±2.08)% in control group (P﹤0.01). Treatment with hispidulin (15 and 30 μmol.L-1) for 24h induced cell cycle arrest at G1/S phase in A-549 cells. Treatment with hispidulin (15、30 and 60μmol.L-1) for 24h in A-549 cells reduced the expression of cyclin D1 and CDK4 proteins and increased levels of p21 protein. The positive expression of proliferation cell nuclear antigen was significantly decreased in A-549 cells treatment with hispidulin (15、30 and 60μmol.L-1) 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 cyclin D1 and CDK4.

Key words: Lung neoplasms, cell line, tumor, cell proliferation, Cell cycle, CyclinDl, cyclin-dependent kinase 4, 周期素依赖激酶抑制剂p21