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目的本研究观察活性维生素D(1,25-dihydroxyvitamin D3,1,25(OH)2D3)和卡铂(carboplatin,CBP)两种药物不同的联合作用方案对SKOV-3细胞增殖的影响,以评价1,25(OH)2D3是否可以增强CBP的抗肿瘤效应。方法本研究分1,25(OH)2D3组,CBP组以及联合作用组。用CCK-8法测定细胞抑制率,使用流式细胞仪进行细胞周期、凋亡、细胞活性氧(Reactive Oxidative Species,ROS)及线粒体膜电位(mitochondrial membrane potential,MMP)等分析。结果1,25(OH)2 D3和CBP单独使用可抑制SKOV-3细胞的增殖,而且联用不同浓度的1,25(OH)2 D3后可以明显降低CBP的半数抑制浓度。实验发现不同的联合用药方案对肿瘤细胞的抑制作用也不同,两种药物同时应用时联合作用系数在0.57～0.78之间,其中10 nmol/L 1,25(OH)2 D3与CBP联用达到高度协同作用。先加1,25(OH)2 D3后加CBP组对SKOV-3细胞的抑制作用明显优于先加CBP后加1,25(OH)2 D3组。细胞周期分析显示,1,25(OH)2 D3使SKOV-3细胞阻滞于G0/G1期,CBP使细胞阻滞于G2/M期,先用1,25(OH)2D3再用CBP使细胞周期发生了双重阻滞作用。1,25(OH)2D3与CBP联用显著提高SKOV-3凋亡率。与正常对照相比较,各处理组均可使细胞ROS的释放增加,MMP下降(P<0.05),其中两种药物同时联合作用方案细胞ROS的产生最多,MMP下降最为显著,其次为先用1,25(OH)2D3后用CBP组。结论 1,25(OH)2D3可以抑制SKOV-3细胞的增殖,并且与CBP同时应用或先于CBP应用时均能显著增强其对卵巢癌细胞的杀伤能力。1,25(OH)2D3与CBP的协同作用与细胞周期的双重阻滞有关,并通过提高ROS和降低MMP抑制卵巢癌细胞的增殖。
Objective To investigate the effects of different combination of 1,25-dihydroxyvitamin D3 (1,25 (OH) 2D3) and carboplatin (CBP) on the proliferation of SKOV-3 cells in order to evaluate 1,25 (OH) 2D3 can enhance the antitumor effect of CBP. Methods This study was divided into 1,25 (OH) 2D3 group, CBP group and combined action group. Cell inhibition rate was measured by CCK-8 method. Cell cycle, apoptosis, reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) were analyzed by flow cytometry. Results 1,25 (OH) 2 D3 and CBP alone could inhibit the proliferation of SKOV-3 cells, and combined with different concentrations of 1,25 (OH) 2 D3 could significantly reduce the half-inhibitory concentration of CBP. The experimental results showed that different combination regimens have different inhibitory effects on tumor cells. The combined effect coefficient of the two drugs was between 0.57 and 0.78. The combination of 10 nmol / L 1,25 (OH) 2 D3 and CBP reached High synergy. After adding 1,25 (OH) 2 D3, the inhibitory effect of CBP group on SKOV-3 cells was significantly better than that of CBP plus 1,25 (OH) 2 D3 group. Cell cycle analysis showed that 1,25 (OH) 2 D3 blocked SKOV-3 cells in G0 / G1 phase and blocked cells in G2 / M phase by CBP. CBP was first used with 1,25 (OH) 2D3 Cell cycle occurred double block effect. 1,25 (OH) 2D3 combined with CBP significantly increased the apoptosis rate of SKOV-3. Compared with the normal control group, the release of ROS increased and the MMP decreased (P <0.05). In the combination of the two drugs, the ROS production was the most and the MMP decreased the most significantly , 25 (OH) 2D3 with CBP group. Conclusion 1,25 (OH) 2D3 can inhibit the proliferation of SKOV-3 cells, and can significantly enhance its cytotoxicity on ovarian cancer cells when applied with CBP or CBP. The synergistic effect of 1,25 (OH) 2D3 and CBP is related to the double arrest of cell cycle, and inhibits the proliferation of ovarian cancer cells by increasing ROS and decreasing MMP.