3-溴丙酮酸联合顺铂抑制肺癌细胞株A549的生长  

Effect of 3-bromopyruvate combined with cisplatin on inhibiting growth of lung cancer A549 cells

作　　者：张梦娇[1] 张明[2] 胡义德[1] 

机构地区：[1]第三军医大学新桥医院肿瘤科,重庆400037 [2]泸州医学院附属医院呼吸内二科,四川泸州646000

出　　处：《第三军医大学学报》2016年第4期338-343,共6页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81372340)~~

摘　　要：目的探讨3-溴丙酮酸(3-bromopyruvate,3-BrPA)联合顺铂体外抗肺癌A549细胞的作用及其可能机制。方法采用CCK-8检测不同浓度的3-BrPA、顺铂单用及联用对A549细胞的增殖抑制;选择低于半数抑制浓度(IC50)的40μmol/L 3-BrPA与1 mg/L顺铂单独和联合作用A549细胞48 h后,用倒置相差显微镜观察细胞形态变化,流式细胞术检测细胞凋亡;不同浓度的3-BrPA作用A549细胞48 h后,流式细胞术检测细胞周期变化,己糖激酶(hexokinase,HK)活性检测试剂盒检测3-BrPA对细胞内HK活性的影响。结果 CCK-8结果显示,与阴性对照组比较,3-BrPA浓度大于20μmol/L时对A549细胞有明显抑制作用(P<0.01),与单用顺铂组比较,3-BrPA联合顺铂组可显著增强顺铂对A549细胞的毒性作用(P<0.01);3-BrPA作用A549细胞48 h后,倒置显微镜观察到3-BrPA组大多数细胞出现凋亡,而联合用药组细胞凋亡更加明显,部分区域出现细胞碎片等坏死征象;流式细胞术结果显示,3-BrPA和顺铂单用组及联合用药组细胞凋亡率均明显高于阴性对照组(P<0.01),且联合用药组细胞凋亡率明显高于单独用药组(P<0.01);细胞周期检测结果显示3-BrPA可将A549细胞阻滞于G1期;HK活性结果显示:与阴性对照组比较,3-BrPA组HK活性明显降低(P<0.01)。结论 3-BrPA有抑制肺癌A549细胞增殖、诱导其凋亡的作用,且与顺铂具有协同抗肺癌A549细胞增殖作用,其机制可能为抑制肺癌细胞糖酵解,进而影响肺癌细胞能量代谢。Objective To in,Jestigate the effect of gIycolydc inhibitor 3-bromopyruvate （5-BrPA） combined with cisplatin on inhibiting lung cancer A549 cells and its possible mechanism. Methods The cell proliferation of A549 cell streated with different concentrations of 3-BrPA and/or cisplatin was detected by CCK-8 assay. 3-BrPA with concentration of 40 ixmol/L, which is lower than the median inhibitory concentration （ICs0）, was selected to combine with cisplatin （1 rag/L） to treat A549 cells. Then, an inverted microscope was used to observe the morphological changes of A549 cells, and flow cytometry was used to detect apoptosis. The A549 cells were treated with different concentrations of 3-BrPA, and then flow cytometry was adopted to detect the changes of cell cycle. The intracellular hexokinase activity was detected with hexokinase activity assay kit. Results Compared with the control group, 3-BrPA with concentration higher than 20 μmol/L showed significantly inhibitory effect on A549 cells （P 〈 0. 01 ）. Compared with cisplatin alone, 3-BrPA in combination with cisplatin significantly enhanced the toxic effect of cisplatin on A549 cells （P 〈 0. 01 ）. After A549 cells were treated with 3-BrPA for 48 h, the morphological changes of cell apoptosis were observed. The combination group showed more obvious morphological changes, and some cells showed signs of necrosis. Flow cytometry indicated that compared with the control group, the apoptotic rates of the 3-BrPA alone group, cisplatin alone group and combination group were significantly higher （ P 〈 O. O1 ）. The apoptotic rates of the 3-BrPA along group and cisplatin alone group were significantly lower than that of the combination group （P 〈 0. 01 ）. The cell cycle assay results showed that 3-BrPA arrested A549 ceils at G1 phase. Hexokinase activity test showed that compared with the control group, the hexokinase activity of 3-BrPA group was significantly lower （P 〈 0. 01 ）. Conclusion 3-BrPA can inhibit the proliferation of A549 c

关 键 词：肺癌 3-溴丙酮酸 顺铂 糖酵解 己糖激酶 

分 类 号：R73-361[医药卫生—肿瘤] R734.2[医药卫生—临床医学]