VX-680联合顺铂对人SK-HEP-1细胞株增殖影响机制探讨

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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 inhibitory effect and chemotherapy sensitivity of Aurora B kinase inhibitor VX-680 combined with cisplatin (DDP) on the proliferation of human hepatoma cell line SK-HEP-1, and to explore its mechanism. METHODS: MTT assay was used to detect the inhibitory effect of VX-680 alone or in combination with DDP on the proliferation of human hepatoma cells SK-HEP-1. Flow cytometry (FCM) was used to analyze the effects of single and combined drugs on the growth, apoptosis and cycle of SK-HEP-1 cells. The expression of Aurora B, p53 and Bcl-2 protein in the experimental group was detected by Western blot. RESULTS: Both VX-680 and DDP inhibited the growth of SK-HEP-1 cells in a dose-dependent manner, and the inhibition rate after combined administration was significantly higher than that of the monotherapy group, P<0.05. When VX-680 was 3.125 μmol/L and DDP was 0.125 μg/mL, a synergistic effect was observed with Q=1.36; FCM showed that the rate of apoptosis was significantly higher in the combination group (17.4±0.3)% than in the control group. The (1.1 ± 0.2)% in the VX-680 group was (5.97 ± 0.5)% and in the DDP group (7.53 ± 0.7)%. The cell cycle results showed that the cells in the combination group decreased in S phase and G1 phase, but the percentage of cells arrested in G2/M phase increased significantly (68.3±1.2)%, which was significantly higher than (16.7±0.9)% and VX in DDP alone. (43.6±1.1)% of -680 and (23.5±0.8)% of control group, P<0.05; and the expression of Aurora B kinase was decreased and the expression of p53 was increased in the combined treatment group by Bcl-2. Reduced expression. CONCLUSION: VX-680 can inhibit the proliferation of SK-HEP-1 cells by promoting apoptosis and effectively increase the chemosensitivity of SK-HEP-1 to DDP. The mechanism is related to the decreased expression of Aurora B, increased expression of p53 and Bcl- 2 related to reduced expression.
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