蛇葡萄素钠对人膀胱癌EJ细胞增殖的抑制作用

Inhibitory Effects of Ampelopsin Sodium on the Proliferation of Human Bladder Cancer Cell Line EJ

目的:研究蛇葡萄素钠(AMP-Na)对人膀胱癌EJ细胞增殖的抑制作用。方法:体外传代培养EJ细胞。以25、33、43.5、57.4、75.8、100μg/ml AMP-Na培养细胞24、48、72 h后,采用MTT法测定细胞活力并计算抑制率。以0、43.5、57.4、75.8μg/ml AMP-Na与2μg/ml顺铂培养细胞24 h后,采用流式细胞仪检测细胞数并计算细胞凋亡率与细胞周期分布。以0、43.5、57.4、75.8μg/ml AMP-Na与2μg/ml顺铂培养细胞24、48 h后,采用实时荧光定量聚合酶链反应(RT-PCR)法测定细胞中Cyclin D1 m RNA的表达。结果:以25、33、43.5、57.4、75.8、100μg/ml AMP-Na培养细胞24、48、72 h后对细胞具有明显抑制作用,且呈明显的时间依赖关系。以57.4、75.8μg/ml AMP-Na培养细胞24、48 h后在G0/G1峰之前出现凋亡峰;以75.8μg/ml AMP-Na培养细胞24 h与43.5、75.8μg/ml培养细胞48 h后,Cyclin D1 m RNA的表达减弱(P<0.05)。结论:AMP-Na能高效抑制EJ细胞的增殖并促进其凋亡,其机制可能与诱导细胞凋亡、下调Cyclin D1 m RNA表达以及使细胞周期阻滞于G0/G1期有关。

OBJECTIVE： To study the inhibitory effects of ampelopsin sodium （AMP-Na） on the proliferation of human blad- der cancer cell line EJ. METHODS： Cell line EJ was subcultured in vitro. MTT assay was adopted to determine cell viability after the cells were cultured in 25μg/ml, 33 μg/ml, 43.5 μg/ml, 57.4μg/ml, 75.8μg/ml, 100μg/ml AMP-Na for 24 h, 48 h and 72 h, and the inhibition rates were calculated by MTT. After the cells were cultured in 0μg/ml, 43.5μg/ml, 57.4μg/ml and 75.8μg/ml AMP-Na and in 2μg/ml cisplatin for 24 h, the flow cytometry was applied to determine the cell quantity, and the apoptosis rates and the percentages of cell-cycle distribution were calculated. After the cells were cultured in the above two kinds of culture solu- tion at the above-mentioned concentrations for 24 h and 48 h, the real-time quantitative fluorescent polymerase chain reaction （RT-PCR） technology was used to determine the expression of Cyclin D1 mRNA in the cells. RESULTS： After the cells were cul- tured in 25μg/ml, 33 μg/ml, 43.5 μg/ml, 57.4 μg/ml, 75.8μg/ml, 100μg/ml AMP-Na for 24 h, 48 h and 72 h, AMP-Na showed obvious inhibitory effects on the cells. The effects was obviously time-dependent. After the cells were cultured in 57.4μg/ml, 75.8μg/ml AMP-Na for 24 h and 48 h, apoptotic peak appeared before GdG1 peak. After the cells were cultured in 75.8 μg/ml AMP-Na for 24 h and in 43.5 μg/ml, 75.8 μg/ml AMP-Na for 48 h, the expression of Cyclin D1 mRNA was decreased （P〈0.05）. CON- CLUSIONS： AMP-Na can effectively inhibit the proliferation of cell line EJ and accelerate its death by a mechanism that may be re- lated to the induction of apoptosis, the reduction of expression of Cyclin D1 mRNA and G0/G1 cell cycle arrest.