论文摘要
目的研究儿茶酚胺类神经递质多巴胺（Dopamine）对人喉癌Hep-2细胞的增殖抑制作用并探讨其可能的作用机制。方法将Hep-2细胞接种于含10％小牛血清、100U/mL双抗（青霉素、链霉素）的DMEM培养基中，37℃、饱和湿度、5％CO2培养箱培养24h，三种不同药物浓度（0.05、0.5、5.0mmol/L）的Dopamine分别处理Hep-2细胞，采用四甲基偶氮唑蓝比色法（MTT法）检测Dopamine对Hep-2细胞增殖活性的影响；流式细胞术（FCM）分析Dopamine对Hep-2细胞周期分布的影响；Hoechst NO.33258荧光染色法荧光倒置显微镜观察Hep-2细胞凋亡形态；酶联免疫吸附测定（ELISA）法检测肿瘤细胞培养上清液中血管内皮生长因子（Vascular endothelial growth factor，VEGF）浓度。结果MTT法检测提示不同浓度的Dopamine作用于Hep-2细胞时，浓度在0.5～5mmol/L时，对Hep-2细胞的生长有明显的抑制作用（P<0.05），但当浓度低于0.5mmol/L时，则无明显抑制作用（P>0.05）。流式细胞术检测结果显示，随着Dopamine浓度的增加，处于G0～G1期细胞的比例逐渐升高（P<0.05），最高浓度时较空白对照组细胞比例升高了（18.28±6.66）％（P<0.05）；S期细胞的比例逐渐降低，最高浓度时较空白对照组细胞比例降低了（17.00±1.23）％（P<0.05）；而G2～M期细胞比例变化则无统计学意义（P>0.05）。ELISA法提示最高浓度（5mmol/L）时VEGF浓度较空白对照组下降了38.5％（P<0.05）。荧光倒置显微镜可观察到染色质边集、核碎裂等典型细胞凋亡形态。结论Dopamine可能通过抑制内生性VEGF的表达对Hep-2细胞的增殖有抑制作用，有望成为治疗喉癌的有效药物。
论文目录
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