天南星多糖对人肾癌细胞系GRC-1增殖及凋亡作用的影响  

Effects of Arisaematis Rhizoma Polysaccharide on Proliferation and Apoptosis of Human Renal Cell Carcinoma Cell Line GRC-1

作　　者：唐化勇[1] 张万生[1] 于航[1] 姜爽[1] 王立国[1] 

机构地区：[1]吉林医药学院附属医院,吉林132000

出　　处：《中国实验方剂学杂志》2016年第14期155-158,共4页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：吉林省教育厅"十二五科学技术研究项目"(吉教科合字2014203);吉林省中医药管理局中医药科技项目(2012163)

摘　　要：目的:探讨天南星多糖(ARPS)对人肾癌细胞系GRC-1的增殖、凋亡及Wnt/β-连接素(β-catenin)信号通路的影响。方法:采用RPMI-1640培养液常规培养获对数生长期的GRC-1细胞后,给予0,20,50,100,200 mg·L^(-1)ARPS处理,0 mg·L^(-1)ARPS组为空白组,细胞计数试剂盒(CCK-8)法检测各浓度ARPS处理24,48,72和96 h的增殖抑制率,分别采用AnnexinFITC/PI双染法和PI单染法检测各浓度处理48 h后的细胞凋亡率和细胞周期情况,蛋白质免疫印迹(Western blot)检测各浓度处理48 h后Wnt/β-catenin信号通路中β-catenin及下游靶分子原癌基因(C-myc),细胞周期素D1(Cyclin D1)蛋白水平。结果:与空白组比较,在20~200 mg·L^(-1),ARPS对GRC-1细胞的增殖有明显抑制效果,细胞增殖抑制率随ARPS作用时间的延长和质量浓度的增加而升高,总体上抑制效应呈剂量和时间依赖的方式,以上差异均有统计学意义(P<0.05),50~200 mg·L^(-1)ARPS的早期凋亡率及20~200 mg·L^(-1)ARPS的晚期凋亡率和总凋亡率均升高(P<0.05),20,50,100,200 mg·L^(-1)ARPS组的G_0期/G_1期高于空白组,S期,G2期/M期及β-catenin,C-myc和Cyclin D1蛋白水平均低于0 mg·L^(-1)ARPS(P<0.05),20~200 mg·L^(-1)各质量浓度间的以上指标的差异均有统计学意义(P<0.05)。结论:ARPS对人肾癌细胞系GRC-1的增殖有一定抑制作用,同时可诱导细胞凋亡和G_0/G_1期阻滞并可抑制Wnt/β-catenin通路激活。Objective： To explore Arisaematis Rhizoma polysaccharide （ARPS） on the proliferation, apoptosis and Wnt/β-catenin signaling pathway of human renal cell carcinoma cell line GRC-1. Method： After entering into the logarithmic growth phase with RPMI-1640 culture fluid under routine condition, the GRC-1 cells were treated with different doses of ARPS （0, 20, 50, 100, 200 mg·L-1 ）. The 0 mg·L-1 ARPS group was the blank group. The inhibition rates of proliferation at 24, 48, 72 and 96 h after the treatment were measured by viable cell counting kit assay （ CCK-8）. The apoptosis rate and cell cycle at 48 h after the treatment were detected by Annexin-FITC/PI double staining and PI staining methods via flow cytometry, respectively. Western blot was employed to detect the expression level of β-catenin in Wnt/β-catenin signaling pathway and its downstream target molecule C-myc and Cyclin D2 at 48 h after the treatment. Result： Compared with the blank group, ARPS within the range of 20-200 mg·L-1 presented an inhibitory effect on GRC-1 cell proliferation. The proliferation inhibition rate increased with the rise in time and concentration of ARPS in a dose-and time-dependent manner, and the above differences were statistically significant （P 〈 0. 05）. Compared with the 0 μg·mL-1 ARPS, the early apoptosis rate of 50-200 mg·L-1 ARPS, the late apoptosis rate of 20-200 mg·L-1 and the total apoptosis rate were all elevated （P 〈0.05）. There was a higher G0/Gl phase ratio than 20, 50, 100, 200 mg·L-1 ARPS groups, and lower S and G2/M phase and β-eatenin, C-myc and Cyclin D1 protein levels than 0 mg·L-1 concentration range between 20-200 mg. L-1 were statistically significant （P （P 〈 0. 05）. Indexes within the 〈 0.05 ）. Conclusion： ARPS inhibited the proliferation of renal cell carcinoma cell line GRC-1, and induced cell apoptosis and G0/G1 arrest, with the effect of inhibiting the activation of Wnt/β-catenin signaling.

关 键 词：天南星多糖 肾癌 抗肿瘤 Wnt/β-连接素(β-catenin)信号通路 

分 类 号：R285.5[医药卫生—中药学]