柴胡皂苷D对胃癌SGC-7901细胞生长、迁移抑制作用和Norrin、Livin的影响

Effect of saikosaponin D on proliferation and migration of human gastric cancer cell line SGC-7901

目的探讨柴胡皂苷D对胃癌SGC-7901细胞生长、迁移抑制作用和Norrin、Livin水平的变化。方法实验分为对照组(正常培养SGC-7901细胞)、低剂量组(SGC-7901细胞+5μmol/L柴胡皂苷D)、中剂量组(SGC-7901细胞+10μmol/L柴胡皂苷D)和高剂量组(SGC-7901细胞+20μmol/L柴胡皂苷D)。采用四甲基偶氮唑蓝(MTT)法检测各组细胞24、48、72h细胞增殖情况;采用TUNEL法检测各组细胞凋亡情况;采用Transwell检测各组细胞迁移能力;采用real-time PCR对各组细胞中Norrin及Livin的表达进行检测;采用Western blot检测各组细胞Norrin、Livin、CDC25A、cyclin A2、p21和cyclin D1表达。结果与对照组比较,低剂量组对SGC-7901细胞生长、凋亡和Norrin、Livin、p21及cyclin D1的表达未产生明显影响(P>0.05),但可减低SGC-7901细胞CDC25A、cyclin A2表达(P<0.05);中、高剂量组SGC-7901细胞生长抑制作用明显,荧光TUNEL显示凋亡率高于对照组(P<0.05),Norrin及Livin mRNA表达减少(P<0.05),CDC25A及cyclin A2表达具有抑制作用(P<0.05),且具有剂量依赖性,对p21及cyclin D1无明显影响(P>0.05)。结论柴胡皂苷D可能通过Norrin、Livin的表达、干扰肿瘤细胞生长周期、介导肿瘤细胞凋亡等作用,抑制人胃癌SGC-7901细胞生长、迁移能力。

Objective To investigate the effect of saikosaponin D on the growth and migration of human gastric cancer cell line SGC-7901.Methods Cultured human gastric cancer SGC-7901 cells were divided into control group and low,medium,high dose groups,which were treated with 0,5,10 and 20 μmol/L saikosaponin D,respectively.The cell proliferation was determined by MTT method at 24 h,48 h and 72 h after treatment;apoptosis was detected by fluorescence TUNEL method,cell migration ability was assayed by Transwell method,the expression of Livin and Norrin mRNA was detected by Real-time PCR,the expression of Norrin,LivinCDC25A,cyclin A2,p21 and cyclin D1 proteins was detected by blot Western blotting.Results Compared to control group,the expression of CDC25A cyclin A2 in low dose group was decreased （P〈0.05）,however,there were no changes in cell apoptosis and expression of Norrin,Livin,p21 and cyclin D1 （P 〉0.05）,in medium and high dose groups,cell proliferation was significantly inhibited,apoptosis rate was increased （P〈0.05）,the expression of Norrin and Livin mRNA was decreased （P〈0.05）,the expression CDC25A and cyclin A2 proteins was inhibited in a dose-effective manner （P〈 0,05）,and there were no changed in expression of p21 and cyclin D1 （P 〉0.05）.Conclusion Saikosaponin D can inhibit the growth and migration of human gastric cancer SGC-7901 cells,which is associated with down-regulating Livin and Norrin expression,interfering with the cell cycle and promoting cell apoptosis.