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目的:研究诺维本(NVB)与热疗联合对人肺癌细胞系A549细胞体外增殖的影响。方法:应用四甲基偶氮唑蓝(MTT)快速比色法测定细胞增殖,确定诺维本的工作浓度,并以该浓度进行化疗或与热疗联合,计算24h细胞生存率,根据Veleriote法判断热化疗联合24h的作用效果;24h流式细胞仪检测A549细胞的凋亡和周期情况。结果:以24h时IC10-IC20的药物浓度作为试验的工作浓度,确定NVB对细胞系的工作浓度为10μg/ml。42℃单独热疗和单独化疗均对该株细胞有抑制和杀伤作用(P<0.05);42℃热疗与药物联合抑制作用强于单独化疗组和单独热疗组(P<0.01);细胞周期分析发现42℃热疗降低了S期细胞比例;单独诺维本使细胞周期阻滞于G2/M期;与单独化疗组相比热化疗组的S期细胞比例减少,G2/M期细胞增多;对照组、热疗组、化疗组、热化疗组细胞凋亡率分别为1.5%、7.9%、13.9%、29.1%。结论:42℃温热可以明显增强化疗药诺维本的毒性,其作用机制可能与干扰细胞周期有关。
OBJECTIVE: To study the effects of combination of NVB and hyperthermia on the proliferation of human lung cancer cell line A549 in vitro. Methods: MTT assay was used to determine cell proliferation. The working concentration of novibenzine was determined. Chemotherapy or hyperthermia was used to calculate the survival rate of 24h cells. According to the Veleriote method To determine the combined effect of thermotherapy and chemotherapy 24h; apoptosis and cycle of A549 cells were detected by flow cytometry 24h. Results: The drug concentration of IC10-IC20 at 24h was used as the working concentration of the test, and the working concentration of NVB on the cell line was determined to be 10 μg / ml. 42 ℃ alone hyperthermia and chemotherapy alone had inhibitory and cytotoxic effects on the cells (P <0.05). Combination of hyperthermia and drug at 42 ℃ had stronger inhibitory effect than chemotherapy alone and hyperthermia alone (P <0.01) Cycle analysis showed 42 ℃ hyperthermia reduced the proportion of S phase cells; Novobib alone block the cell cycle in G2 / M phase; Compared with chemotherapy alone group S phase cells decreased, G2 / M phase cells The apoptosis rates of control group, hyperthermia group, chemotherapy group and thermochemotherapy group were 1.5%, 7.9%, 13.9% and 29.1% respectively. CONCLUSION: Warming at 42 ℃ can significantly enhance the toxicity of chemotherapy drug Novubibrio, and its mechanism may be related to interfering with the cell cycle.