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目的观察活性维生素D[1,25-dihydroxyvitamin D3,1,25(OH)2D3]和卡铂(carboplatin,CBP)两种药物单独及联合作用对人肺癌A549细胞的抗肿瘤效应,并探讨其可能的作用机制。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,使用激光扫描共聚焦显微镜进行维生素D受体(vitamin D receptor,VDR)的鉴定,用流式细胞仪进行细胞周期、细胞活性氧(reactiveoxidative species,ROS)及线粒体膜电位(mitochondrial membrane potential,MMP)的分析。结果 1,25(OH)2D3和CBP单独使用均可以抑制A549细胞的生长,且两者联合作用时具有协同作用,1,25(OH)2D3能显著提高CBP对A549细胞的抑制率。通过免疫荧光测得A549细胞中有较高维生素D受体的表达。细胞周期分析显示,经1,25(OH)2D3和CBP处理后的A549细胞,细胞周期发生变化,G0/G1期细胞数增多,S期和G2/M期细胞相应减少。两者联合作用后,以上趋势更加明显。与正常对照组相比较,各处理组均可使细胞ROS的释放增加(P<0.05),MMP下降(P<0.05),其中两种药物联合作用时细胞ROS的产生最多,MMP下降最为显著。结论 1,25(OH)2D3可以抑制A549细胞的增殖,并且与CBP联合作用时能显著增强其对肺癌细胞的杀伤能力。1,25(OH)2D3与CBP的协同作用与阻滞细胞周期的有序进行有关,并通过提高ROS和降低MMP抑制肺癌细胞的增殖。
Objective To investigate the antitumor effects of two drugs 1,25-dihydroxyvitamin D3, 1,25 (OH) 2D3 and carboplatin (CBP) alone and in combination on human lung cancer A549 cells and to explore their possible effects The mechanism of action. Methods This study was divided into 1,25 (OH) 2D3 group, CBP group and combined action group. Cell inhibition rates were determined by CCK-8 assay, and vitamin D receptor (VDR) was identified by laser scanning confocal microscopy. Cell cycle, reactive oxygen species (ROS) and Analysis of mitochondrial membrane potential (MMP). Results 1,25 (OH) 2D3 and CBP alone could inhibit the growth of A549 cells. Synergistic effect was observed when combined with 1,25 (OH) 2D3 and 1,25 (OH) 2D3 could significantly increase the inhibitory rate of CBP on A549 cells. A549 cells were assayed for higher vitamin D receptor expression by immunofluorescence. Cell cycle analysis showed that A549 cells treated with 1,25 (OH) 2D3 and CBP changed the cell cycle, increased the number of cells in G0 / G1 phase and decreased the cells in S phase and G2 / M phase. After the combined effect of the two, the above trend is even more obvious. Compared with the normal control group, the release of ROS increased (P <0.05) and the MMP decreased (P <0.05). The combination of the two drugs produced the most ROS and the most significant decrease of MMP. Conclusions 1,25 (OH) 2D3 can inhibit the proliferation of A549 cells, and when combined with CBP, it can significantly enhance its killing ability on lung cancer cells. The synergistic effect of 1,25 (OH) 2D3 and CBP is related to the block of the orderly progression of cell cycle, and inhibits the proliferation of lung cancer cells by increasing ROS and decreasing MMP.