(R,R')-富马酸戊乙奎醚通过诱导细胞周期G_0/G_1期阻滞抑制非小细胞肺癌H157细胞增殖  

(R,R')-penehyclidine fumarate inhibits non-small cell lung cancer H157 cell growth and arrests the cell cycle in G_0/G_1

作　　者：华南[1] 魏晓莉[1] 何新华[1] 仲伯华[1] 郑建全[1] 

机构地区：[1]军事医学科学院毒物药物研究所,北京100850

出　　处：《中国新药杂志》2012年第5期543-547,554,共6页Chinese Journal of New Drugs

摘　　要：目的:考察抗胆碱药(R,R')-富马酸戊乙奎醚[(R,R')-penehyclidine fumarate,R2PHF]对非小细胞肺癌H157细胞增殖的影响,探讨其作用机制。方法:应用RT-PCR检测H157上M受体的表达;磺酰罗丹明B(sulforhodamine B,SRB)法测定R2PHF作用于H157后细胞活力改变;流式细胞术(FCM)检测细胞周期和细胞凋亡;Western Blot分析细胞周期蛋白cyclin D1表达水平的变化。结果:M受体五种亚型在H157上有不同的表达,R2PHF在体外可时间及浓度依赖性抑制H157细胞增殖并诱导其发生G0/G1期阻滞。FCM检测结果显示,20μmol.L-1R2PHF作用48 h后,H157细胞G0/G1期比率显著增高,S期比率显著下降,而细胞凋亡未发生明显变化。Western Blot分析表明R2PHF可时间及浓度依赖性下调细胞周期蛋白cyclin D1的表达。结论:R2PHF抑制非小细胞肺癌H157细胞增殖与其下调cyclin D1的表达水平并将细胞阻滞在G0/G1期密切相关。Objective： To investigate the effect of cholinergic antagonist（R,R＇）-penehyclidine fumarate（R2PHF） on proliferation of human non-small cell lung cancer H157 cells and explore the potential mechanism.Methods： The muscarinic receptors expressed in H157 cells were detected by RT-PCR.H157 cells were treated with different concentration of R2PHF for various time,and cell viability was detected by sulforhodamine B（SRB）.Flow cytometry（FCM） was used to analyze cell cycle and apoptosis after treatment with R2PHF.The expression of cyclin D1 was analyzed by Western Blot.Results： Expressions of the five muscarinic receptor subtypes were different in H157 cells.The proliferation of H157 cells was inhibited by R2PHF in a concentration-and time-dependent manner in vitro and the cell cycle was arrested in G0/G1.The percentage of G0/G1 phase cells increased while the S phase cells decreased after treated with 20 μmol·L-1 R2PHF for 48 h.There was no change observed in cell apoptosis.R2PHF significantly down regulated the expression of cyclin D1 in a concentration-and time-dependent manner.Conclusion： These results reveal that R2PHF could inhibit proliferation of non-small cell lung cancer H157 cells in vitro by down-regulating expression of cyclin D1 and arresting the cell cycle in G0/G1 phase.

关 键 词：(R R')-富马酸戊乙奎醚 非小细胞肺癌 抗胆碱药 细胞增殖 细胞周期 

分 类 号：R979.1[医药卫生—药品]