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目的:研究VP-16与热疗联合对人肺癌细胞系SBC5细胞株体外增殖的影响。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VP-16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热疗作用效果;24h流式细胞仪检测SBC5细胞的凋亡和周期情况;电镜观察不同处理后细胞超微结构。结果:以24hIC10-IC20的药物浓度作为试验的工作浓度,确定VP-16对细胞的工作浓度为1μg/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期细胞所占比例;单独VP-16使细胞周期阻滞于G2/M期;与单独化疗组相比热化疗组的G2/M期细胞所占比例减少,S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3.3、6.9、10.3、21.1;透射电镜下见热疗后细胞胞膜起泡,胞浆浓缩,细胞器完整,核染色质浓集于胞核周边。结论:42℃温热可以明显增强化疗药VP-16的毒性,其作用机制可能与干扰细胞周期和改变细胞超微结构有关。
Objective: To investigate the effects of VP-16 combined with hyperthermia on proliferation of human lung cancer cell line SBC5 in vitro. Methods: MTT assay was used to determine cell proliferation. The working concentration of VP-16 was determined. Chemotherapy or hyperthermia was used to calculate the cell survival rate of 24h. According to the Veleriote method The effect of hyperthermia combined with 24h hyperthermia was evaluated. The apoptosis and cycle of SBC5 cells were detected by flow cytometry in 24 h. The ultrastructures of SBC5 cells were observed by electron microscopy. Results: The drug concentration of 24hIC10-IC20 was used as the experimental working concentration, and the working concentration of VP-16 on the cells was determined to be 1 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 that 42 ℃ hyperthermia reduced the proportion of S phase cells; VP-16 alone blocked cell cycle in G2 / M phase; compared with chemotherapy alone group, G2 / M phase cells decreased proportion of cells , S phase cells increased; the control group, hyperthermia group, chemotherapy group, thermochemotherapy group apoptosis rates were 3.3,6.9,10.3,21.1; under the transmission electron microscope after hyperthermia cell membrane blistering, cytoplasm concentration, Organelles complete, nuclear chromatin concentrated in the peribronchia. CONCLUSION: Warming at 42 ℃ can significantly enhance the toxicity of chemotherapeutic drug VP-16. The mechanism may be related to interfering with cell cycle and changing cell ultrastructure.