摘要
目的检测鞣花酸（EA）对胶质瘤U251细胞增殖、侵袭能力的影响并探讨其机制。方法利用细胞计数试剂盒（CCK-8）法检测EA对U251细胞增殖能力的影响；利用流式细胞术检测EA对U251细胞凋亡和周期的影响。利用划痕实验、Transwell侵袭实验检测EA对U251细胞侵袭的影响。应用Western blot检测EA对U251细胞中B细胞淋巴瘤/白血病-2（bcl-2）、B细胞淋巴瘤/白血病-2相关X蛋白（bax）、半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-3、基质金属蛋白酶（MMP）-9、MMP-2表达水平的影响。结果EA能够明显抑制U251细胞的增殖，抑制作用呈时间、浓度依赖性，处理细胞48 h的半数抑制（IC50）浓度为39.24 μg/ml；流式凋亡分析中对照组、40 μg/ml的凋亡率分别为（2.41±1.74）%、（34.07±0.28）%，差异有统计学意义（P＝0.001），表明EA能促进U251细胞凋亡。流式周期分析中对照组、40μg/ml组S期细胞百分比分别为（20.45±0.28）%、（47.75±2.02）%，差异有统计学意义（P＝0.000），表明EA能引起细胞周期S期阻滞。EA作用48 h后，U251细胞中bax、Caspase-3的表达水平升高，bcl-2的表达水平下降。对照组、EA 5 μg/ml和10 μg/ml组24 h的划痕愈合率分别为（53.88±0.30）%、（36.82±0.56）%、（31.33±0.24）%，差异有统计学意义（P＝0.000，P＝0.000）；Transwell侵袭实验中对照组、EA 5 μg/ml和10 μg/ml组24 h的穿膜细胞数分别为（168.2±7.1）、（66.4±2.5）、（38.6±1.5）个，差异有统计学意义（P＝0.001，P＝0.000）。EA作用48 h后，U251细胞中MMP-9、MMP-2的表达水平下降。
结论EA能够抑制脑胶质瘤细胞U251的增殖和侵袭能力。
Objective To investigate the anti-proliferation and anti-metastatic effect of ellagic acid （EA） on glioma cell line U251 and related mechanisms.Methods In this study, we employed cell counting kit-8 （CCK-8） assay to test the anti-proliferative effect of EA. Flow cytometer was used to monitor cell apoptosis and cell cycle. Wound healing assay and Transwell assay were used to test the anti-invasion effect of EA. Western blotting was employed to test the effect of EA on the expressions of B cell lymphoma/leukemia-2 （bcl-2）, bcl-2 associated X protein （bax）, Cysteinyl aspartate-specific protease （Caspase）-3, matrix metalloproteinase （MMP）-9 and MMP-2.Results EA could significantly inhibit the proliferation of U251 cells in concentration-dependent manner and time-dependent manner, and the concentration of half maximal inhibitory concentration （IC50） after treated 48 hours was 39.24 μg/ml. The apoptosis rate of 40 μg/ml was （34.07±0.28）%, compared with control group, the difference was significant （P=0.001）. Flow cytometer test shows the percentage of cells in S phase treated by EA 40 μg/ml was （47.75±2.02）%, compared with control group （20.45±0.28）%, the difference was significant （P=0.000）. The expressions of bax and Caspase-3 were increased significantly, while bcl-2 was decreased. In Wound healing assay, when treated 24 h by EA, the wound healing rate in control group, 5 μg/ml group, 10 μg/ml group were （53.88±0.30）%, （36.82±0.56）% and （31.33±0.24）%, the difference was significant （P=0.000, P=0.000）; In Transwell assay, the number of passed cells were （168.2±7.1）, （66.4±2.5） and （38.6±1.5） respectively, the difference was significant （P=0.001, P=0.000）. The expressions of MMP-9 and MMP-2 were decreased significantly.Conclusion EA inhibits the proliferation and invasion of glioma cell line U251, which suggests that EA is a potential therapeutic agent for glioma.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2018年第3期463-466,共4页
Chinese Journal of Experimental Surgery
