华蟾素联合三氧化二砷对结肠癌细胞的作用

Effect of cinobufagin combined with As_2O_3 on colon cancer cells

目的探讨华蟾素联合三氧化二砷(As_2O_3)对结肠癌细胞的作用。方法将结肠癌细胞株SW620分为四组:As_2O_3组中分别加入As_2O_30.10ng/ml(A1组)、0.25ng/ml(A2组)、0.50ng/ml(A3组)、1.00ng/ml(A4组)、2.00ng/ml(A5组)(每个浓度6个复孔);华蟾素组中分别加入华蟾素5μg/ml(H1组)、10μg/ml(H2组)、15μg/ml(H3组)、20μg/ml(H4组)、25μg/ml(H5组);联合组中分别加入As_2O_30.1ng/ml+华蟾素10μg/ml(L1组)、As_2O_30.5ng/ml+华蟾素10μg/ml(L2组)、As_2O_32.0ng/ml+华蟾素10μg/ml(L3组);取正常培养的细胞作为对照组。比较各组细胞增殖、周期、凋亡及相关蛋白的表达。结果细胞增殖抑制率随着华蟾素和As_2O_3浓度的增加而增加(P<0.05),联合用华蟾素和As_2O_3后,细胞增殖抑制率增强,且随着As_2O_3浓度的增加而增加(P<0.05)。与对照组比较,L1组、L2组、L3组G_0/G_1期细胞比例及细胞凋亡增加(P<0.05),且随着As_2O_3浓度升高而增加(P<0.05)。与对照组、H2组比较,L1组、L2组、L3组p21WAF1、p27和Bax蛋白表达增加(P<0.05或P<0.01),并随着As_2O_3浓度升高而增加(P<0.05或P<0.01),Bcl-2蛋白表达减少,并随着As_2O_3浓度的增加而减少(P<0.01)。结论华蟾素联合As_2O_3能抑制结肠癌细胞的生长,具有协同抗肿瘤效应,可能与p21WAF1、p27和Bax蛋白表达上调、Bcl-2蛋白表达下调有关。

Objective To investigate the effect of cinobufagin combined with arsenic trioxide（As2O3）on colon cancer cells.Methods The colon cancer cell line SW620 was divided into four groups.Group A was treated with As2O3 0.10ng/ml（group A1）,0.25ng/ml（group A2）,0.50ng/ml（group A3）,1.00ng/ml（group A4）and 2.00ng/ml（group A5）.Group H was treated with cinobufagin 5μg/ml（group H1）,10μg/ml（group H2）,15μg/ml（group H3）,20μg/ml（group H4）and 25μg/ml（group H5）.Group L was treated with As2O3 0.1ng/ml＋cinobufagin 10μg/ml（group L1）,As2O3 0.5ng/ml＋cinobufagin 10μg/ml（group L2）and As2O3 2.0ng/ml＋cinobufagin 10μg/ml（group L3）.Normal cells were taken as the controls（group C）.The ability of proliferation,cell cycle,cell apoptosis and protein expressions were compared.Results The inhibition rate of cell proliferation was increased as the increase of the concentration of cinobufagin and As2O3（P〈0.05）.The cinobufagin combined with As2O3 promoted the inhibition of cell proliferation,which was increased as the increase of the concentration of As2O3（P〈0.05）.The proportion of G0/G1 and cell apoptosis were higher in groups of L1,L2 and L3 than those in group C,which were rised as the increase of the concentration of As2O3（P〈0.05）.Compared to groups of C and H2,the protein levels of p21WAF1,p27 and Bax were higher and Bcl-2 was lower in groups of L1,L2 and L3,which were aggravated as the increase of the concentration of As2O3（P〈0.05 or P〈0.01）.Conclusion The cinobufagin combined with As2O3 has a synergistic inhibitory effect on the growth of colon cancer cell,which may be related to the upregulation of p21WAF1,p27 and Bax and the downreguation of Bcl-2.