辛伐他汀联合顺铂对人非小细胞肺癌A549细胞增殖的抑制作用

Inhibition of simvastatin combined with cisplatin on human non-small cell lung carcinoma A549 cells

目的研究辛伐他汀联合顺铂对人非小细胞肺癌A549细胞的抑制作用,并探讨其作用机制。方法采用MTT法检测0.625、1.25、2.5、5、10μmol/L顺铂和3.125、6.25、12.5、25、50μmol/L辛伐他汀对A549细胞增殖抑制率的影响;考察2.5μmol/L顺铂与3.125、6.25、12.5、25、50μmol/L辛伐他汀联用对A549细胞增殖抑制作用的协同效果;Annexin V-FITC/PI双染后流式细胞术检测5μmol/L顺铂与25μmol/L辛伐他汀联合应用对对A549细胞凋亡率的影响;分光光度法检测辛伐他汀(25μmol/L)联合顺铂(2.5μmol/L)对A549细胞caspase-3活性的影响。结果辛伐他汀或顺铂对A549细胞的增殖有显著的抑制作用,随着浓度的增加、时间的延长,呈时间、剂量相关性,其抑制作用增强(P<0.01)。顺铂(2.5μmol/L)与不同浓度(3.125、6.25、12.5、25、50μmol/L)辛伐他汀联用对A549细胞的增殖有抑制作用,随着时间的延长,辛伐他汀剂量的增加,两者联用抑制作用呈增强的趋势(P<0.01),2.5μmol/L顺铂联用辛伐他汀25μmol/L的抑制作用具有协同效果。25μmol/L辛伐他汀和2.5μmol/L顺铂能有效地诱导A549细胞的凋亡,而且两者联合使用可以显著增加A549细胞的凋亡率。25μmol/L辛伐他汀联合2.5μmol/L顺铂应可以显著增加A549细胞的caspase-3酶活性。结论辛伐他汀能够显著增强顺铂对A549细胞增殖抑制、诱导凋亡的作用,可能通过上调caspase-3活性诱导A549细胞凋亡。

Objective To evaluate inhibitory effect of simvastatin combine with cisplatin onnon-small cell lung carcinoma A549 cell, and explore its mechanism. Methods Effects of 0.625, 1.25, 2.5, 5, and 10 μmol/L cisplatin and 3.125, 6.25, 12.5, 25, and 50 μmol/L simvastatin on the proliferation inhibition rate of A549 cells were determined by MTT method. Synergistic effects of 2.5 μmol/L cisplatin combined with 3.125, 6.25, 12.5, 25, and 50 μmol/L simvastatin on the proliferation inhibition of A549 cells were studied. Annexin V-FITC/PI double staining flow cytometry was used to detect the effect of 5 μmol/L cisplatin combined with 25 μmol/L simvastatin on the apoptosis rates of A549 cells. Effects of simvastatin（25 μmol/L） combined with cisplatin（2.5 μmol/L） on the caspase-3 activity of A549 cells were measured by spectrophotometry. Results Simvastatin or cisplatin could significantly inhibit the proliferation of A549 cells in a concentration-and time-dependent manner. With the increase of concentration and the prolongation of time, the inhibitory effect was enhanced（P〈0.01）. Cisplatin（2.5 μmol/L） combined with simvastatin with different concentrations（3.125, 6.25, 12.5, 25, and 50 μmol/L） had inhibition on proliferation of A549 cells. With time extension and simvastatin increase, the inhibition was enhanced with the trend of combination（P〈0.01）. Cisplatin（2.5 μmol/L） combined with simvastatin（25 μmol/L） had synergistic effect on inhibitory effects. Simvastatin（25 μmol/L） and cisplatin（2.5 μmol/L） could effectively induce apoptosis of A549 cells, and the combination could significantly increase the apoptosis rate of A549 cells. The caspase-3 activity of A549 cells could be significantly enhanced treated by simvastatin（25 μmol/L） and cisplatin（2.5 μmol/L）. Conclusion Simvastatin can significantly enhance the proliferation inhibition and apoptosis induced by cisplatin in A549 cells, and it may induce apoptosis of A549 cells by up regulation of caspase-3 activity.