VX-680联合顺铂对人SK-HEP-1细胞株增殖影响机制探讨

Effect of VX-680 combined with DDP on proliferation in hepatoma SK-HEP-1 cells and its mechanism

目的:体外观察Aurora B激酶抑制剂VX-680与顺铂(cisplatin,DDP)联合对人肝癌细胞SK-HEP-1细胞株增殖的抑制作用及化疗敏感性,并且初步探讨其机制。方法:MTT法检测VX-680单独或联合DDP对人肝癌细胞SK-HEP-1增殖的抑制作用。流式细胞术(flow cytometry,FCM)分析单独用药及联合用药对SK-HEP-1细胞生长凋亡和周期的影响。蛋白质印迹法检测实验组中Aurora B、p53和Bcl-2蛋白的表达。结果:VX-680与DDP均能抑制SK-HEP-1细胞的生长,呈剂量依赖性,并且联合用药后的抑制率明显高于单用药组,P<0.05。当VX-680为3.125μmol/L和DDP为0.125μg/mL联合时,产生协同作用,Q=1.36;FCM检测发现,联合用药组(17.4±0.3)%的细胞凋亡率明显高于对照组的(1.1±0.2)%,VX-680组为(5.97±0.5)%,DDP组为(7.53±0.7)%。细胞周期结果显示,联合用药组S期和G1期细胞减少,而停滞在G2/M期细胞比例明显增加(68.3±1.2)%,明显高于单用药组DDP的(16.7±0.9)%和VX-680的(43.6±1.1)%及对照组的(23.5±0.8)%,P<0.05;并且通过蛋白质印迹检测发现,在联合用药组中的Aurora B激酶表达减少,p53表达增加,Bcl-2表达减少。结论:VX-680能够通过促进细胞凋亡抑制SK-HEP-1细胞的增殖,有效提高SK-HEP-1对DDP的化疗敏感性,其机制与Aurora B的表达减少、p53表达增加及Bcl-2表达减少有关。

OBJECTIVE：To investigate the effects of an Aurora kinase B inhibitor （VX-680） combined with cisplatin on the growth of liver cancer SK-HEP-1 cells and study the initial mechanism. METHODS： MTT assay was used to exam- ine the inhibition rate of cell growth when cells were treated at various concentrations of VX-680 and DDP alone or combi- nation. Cell cycle and apoptosis rate of SK-HEP-1 cells were detected by flow cytometry after using drugs. The Aurora B, p53,Bcl-2 protein were detected by Western Blot. RESULTS： The growth of SK-HEP-1 cells treated with VX-680, DDP and the combination of both was inhibited in dose-dependent manners. The inhibition rate of the combined treatment was significantly higher than that of two other single drug groups （P〈0.05）. VX-680 in the concentration of 3. 125 μmol/L and DDP in the concentration of 0. 125 μg/mL interaction had the synergistic effect （Q= 1.36）. The result of FCM showed that the apoptosis rate of combined group （5.97±0.5） % was higher than that of the control group （1.1±0.2） % or single group [VX-680 （5. 97±-0. 5）% DDP （7. 53 ± 0. 7）%]. The cell cycle analysis indicated that the rate of SK-HEP-1 cells reduced in G0/G1 and S phase. The rate of SK-HEP-1 cells in Gz phase （68.3 ± 1.2） % were much higher than the single drug group [DDP （16.7±0. 9）% ;VX-680 （43.6±1.1）%] and control group （23.5±0.8）% （P〈 0.05）. The result of Western blot showed that VX-680 significantly inhibited the expression of Aurora-B. VX-680, DDP and the combination of both reduced the expression of BCL-2 protein, raised p53 protein expression, whose effects were more obvious in the combined therapy. CONCLUSIONS： VX-680 could effectively induce SK-HEP-1 chemosensitivity toDDP. Its mechanism was regulated.