不同温度下紫杉醇对人肺癌A549细胞抑制效应
Inhibition effect of paclitaxel on human carcinoma A549 cells at different temperature
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摘要:目的 研究不同温度下紫杉醇对人肺癌A549细胞的增殖抑制效应及对血管内皮生长因子mRNA(VEGF mRNA)表达的影响。方法 采用不同的温度(37, 39, 41, 43和45℃)及不同剂量的紫杉醇(6.25, 12.5, 25, 50, 100和200ng/ml), 并采用组内分组设计, 共30个组。应用四甲基偶氮噻唑蓝(MTT)比色法测定热疗、化疗及热化疗联合对A549细胞的抑制率; 以β-actin基因为参照, 应用半定量RT-PCR技术检测不同剂量紫杉醇及其与加热联合对A549细胞VEGF mRNA表达的影响。结果 43℃组细胞抑制率与不同温度同剂量组间细胞抑制率比较, 除与41℃50 ng/ml和45℃100 ng/ml组对比差异无统计学意义外(P > 0.05), 其余各组差异均有统计学意义(P < 0.05);39℃加热组各个剂量平均抑制率均低于37℃组, 200 ng/ml剂量组间的差异有统计学意义(P < 0.01);43℃加热40 min后的0, 6.25, 12.5, 25 ng/ml剂量组的VEGF mRNA表达水平均低于37℃, 0和6.25 ng/ml组差异有统计学意义(P < 0.01)。结论 39℃加热40 min可能会降低紫杉醇对A549细胞的抑制效应; 43℃加热40 min可以增加A549细胞对紫杉醇的敏感性; 紫杉醇可以降低A549细胞VEGF mRNA的表达; 43℃加热40 min可以增加低剂量紫杉醇对VEGF mRNA表达的抑制效应。Abstract:Objective To investigate the effects of hyperthermia on human carcinoma A549 cell in proliferation and Vascular Endothelial Growth Factor(VEGF)mRNA expression with Paclitaxel and to provide theo retic basis for clinical practices.Methods thirty groups were divided and different temperatures(39℃, 41℃, 43℃ and 45℃)of hyperthermia and different concentrations(6.25, 12.5, 25, 50, 100 and 200 ng/ml)of Paclitaxel were administrated methyl thiazolyl tetr azolium(MTT)assay was adopted to measure the inhibiting rates on cell growth.Semi-quantitative RT-PCR was applied to determine the expression level of VEGF mRNA.Results the inhibiting rates at 43℃ were hig her than those at other temper atures at the same doses significantly(P < 0.05)except the groups of 41℃ 50 ng/ml and 45℃ 100 ng/ml.the inhibiting rates at 39℃ were lower than other groups at the same doses.VEGF mRNA expression levels of the groups at 0, 6.25, 12.5 and 25 ng/ml became lower after hyper thermia for 40min at 43℃ and the differences at the doses of 0 and 6.25 ng/ml were significant(P < 0.01)Conclusion ther motherapy for 40 min at 43℃ would increase the sensitivity, but decrease the sensitivity at 39℃ to Paclitaxel.Paclitaxel could suppress the expression of VEGF mRN A.thermotherapy for 40min at 43℃ would enhance the inhibiting effects on VEGF mRNA expression at the lower doses of Paclitaxel.
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