Zebularine联合丁酸钠对结肠癌LOVO细胞及Runx3、P21的影响

摘要

目的 探讨Zebularine[1-(β-D-呋喃核糖苷)-1,2-二氢嘧啶-2-酮]和丁酸钠(sodium butyrate)对人结肠癌LOVO细胞Runx3及P21的影响.方法 人结肠癌细胞系LOVO按常规培养于含100g/L胎牛血清的PRMI 1640培养液中,分为4组:对照组、Zebularine组、丁酸钠组以及联合用药组,四甲基偶氮唑蓝(MTT)法检测细胞处理24、48、72h后的细胞抑制率,Annexin V-FITC/PI双染法检测细胞凋亡,RT-PCR检测Runx3 mRNA、P21mRNA的表达.结果 单独应用Zebularine及丁酸钠72h后,细胞生长抑制率及凋亡率与对照组相比增高,联合用药较单独用药抑制率及凋亡率明显增高(P＜0.05),生长抑制率呈时间及浓度依赖性,单独用药组Runx3、P21 mRNA的表达相对增加,联合用药组其表达明显增加,与单用药组相比,差异有统计学意义(P＜0.05)；结论 与单用Zebularine及丁酸钠相比,联合用药更显著增加Runx3 mRNA、P21 mRNA的表达,抑制细胞增殖及促进凋亡.%Objective To explore the effects of Zebularine and sodium butyrate on RUNX3 and P21 in LOVO cells.Methods LOVO cells were cultured in RPMI-1640 culture medium containing 100g/L fetal calf serum,the trial consisted of four groups:control group,Zebularine group,sodium butyrate group and Zebularine + sodium butyrate group.The cell viability was examined by an MTT assay and apoptosis was examined by Annexin V-FITC/PI double staining.The mRNA expression of P21 and RUNX3 was determined by reverse transcriptase polymerase chain reaction (RT-PCR).Results The cell growth inhibition rate and apoptosis rate increased in Zebularine group and sodium butyrate group (P ＜ 0.05),and significantly increased in combination group compared with the zebularne group and Sodium butyrate group (P ＜ 0.05).The cell growth inhibition was in a time-and dose-dependent pattern.The relative expression of RUNX3 mRNA and P21 mRNA treated with Zebularine and sodium butyrate was increased compared with the control.It was significantly increased in combination group,and there were significant differences between combination gronp and single drug group (P ＜0.05).Conclusion Compared with zebularine or Sodium butyrate alone,combination of two drugs can significantly induce P21 mRNA and RUNX3 mRNA expression and inhibit the proliferation and facilitate the apoptosis of cell line LOVO.