新藤黄酸体外抑制肺腺癌A549细胞增殖的作用

Inhibitive Effects of Gambogenic Acid on Proliferation of Lung Adenocarcinoma A549 Cells In Vitro

目的研究新藤黄酸（Gambogenic acid,GNA）抑制肺腺癌细胞株A549增殖的作用及其相关机制。方法不同浓度GNA分别作用A549细胞24、48和72 h,甲基噻唑基四唑（methyl thiazolyl tetrazolium,MTT）比色法检测药物对A549细胞增殖的抑制率,吖啶橙/溴化乙啶（acridine orange/ethidium bromide,AO/EB）染色荧光显微镜检测细胞的凋亡;乳酸脱氢酶（lactate dehydrogenase,LDH）法检测GNA对A549细胞损伤作用。结果 MTT法检测结果显示,GNA在0-8μmol/L浓度范围内、24-72 h时间范围内对A549的生长有明显抑制作用,且呈浓度和时间依赖关系;相差显微镜观察表明GNA作用A549细胞48 h后,细胞变圆,呈半贴壁状态;AO/EB染色荧光显微镜观察显示GNA作用48 h后,细胞出现典型的凋亡特征;LDH实验进一步证实GNA在0-4μmol/L浓度范围内对正常细胞未见明显毒性作用。与空白对照组比较,8μmol/L GNA显著降低LDH释放。结论 GNA具有体外抑制肺腺癌细胞株A549增殖的作用,在0-4μmol/L浓度范围内GNA可能是通过诱导细胞凋亡而发挥抑制A549细胞增殖的作用。

Objective To investigate anti-proliferation effect of Gambogenic acid（GNA） on lung adenocarcinoma A549 cells and its underlying mechanisms.Methods A549 cells were treated with different concentration of GNA for 24,48 and 72 h,respectively;the inhibitive effect was measured by methyl thiazolyl tetrazolium（MTT） assay;alternation of ultra structure was detected by AO/EB staining under fluorescent microscope;lactate dehydrogenase（LDH） assay was applied to determine whether GNA induced A549 cells necrosis.Results MTT assay evidenced that GNA inhibited A549 cells proliferation in a dosage dependent manner within the concentration ranging from 0 to 8 μmol/L and time-dependent manner from 24 to 72 h;AO/EB staining under fluorescent microscope showed that 24 h GNA treatment could induce typical apoptosis in A549 cells;LDH assay showed that no significant difference in the LDH release was observed after GNA treatment for 24 h in the concentration ranging from 1 to 4 μmol/L,while 8 μmol/L GNA could statistically induce LDH release in comparison with the control group（P〈0.01）.Conclusion GNA has the capacity to inhibit the proliferation of human lung adenocarcinoma A549 cells.Its underlying mechanisms is related to apoptosis inducing.