β-桐酸对人绒毛膜上皮癌JEG-3细胞凋亡的影响

Effect ofβ-Eleostearic Acid on the Apoptosis of Human Choriocarcinoma JEG-3 Cells

目的:研究β-桐酸对人绒毛膜上皮癌JEG-3细胞凋亡的影响,初步探讨其作用机制。方法:以不同浓度的β-桐酸处理体外培养的绒癌JEG-3细胞,应用MTT实验检测细胞相对活力;应用乳酸脱氢酶(lactate dehydrogenase,LDH)释放法检测β-桐酸对JEG-3细胞的毒性作用;通过流式细胞术和蛋白免疫印迹法分析细胞凋亡的变化;活性氧(reactive oxygen species,ROS)水平通过对2',7'-二氯荧光素(2',7'-dichlorofluorescein,DCF)荧光强度的检测确定;应用ROS抑制剂N-乙酰-L-半胱氨酸(N-Acetyl-L-Cysteine,NAC)分析ROS在β-桐酸对JEG-3细胞活力影响中的作用。结果:β-桐酸对JEG-3细胞活力有明显的抑制作用,且呈剂量和时间依赖性(P<0.05);β-桐酸显著促进LDH的释放和JEG-3细胞的凋亡(P<0.05),凋亡相关蛋白Caspase-3的表达水平与Bax/Bcl-2的表达比例均上调,进一步证明β-桐酸能够诱导JEG-3细胞凋亡;β-桐酸处理后,JEG-3细胞内ROS的含量上升,NAC能够显著拮抗β-桐酸对JEG-3细胞活力的抑制作用(P<0.05)。结论:β-桐酸能够诱导JEG-3细胞的凋亡,其作用可能与ROS的产生有关。

Objective:To study the effect ofβ-Eleostearic acid on the apoptosis of human choriocarcinoma JEG-3 cells and its relative mechanisms.Methods:JEG-3 cells were treated with different concentrations ofβ-eleostearic acid.MTT was used to detect the relative activity of cells.The release of lactate dehydrogenase(LDH)was detected according to the instructions to analyze the toxic effect ofβ-eleostearic acid on JEG-3 cells.Flow cytometry and western blotting were used to analyze the apoptosis of JEG-3 cells.The reactive oxygen species(ROS)level was determined by the fluorescence intensity of 2',7'-dichlorofluorescein(DCF).ROS inhibitor N-acetyl-L-cysteine(NAC)was used to analyze the role of ROS in the effect ofβ-eleostearic acid on the relative activity of JEG-3 cells.Results:β-eleostearic acid could inhibit the relative activity of JEG-3 cells in a dose-dependent and time-dependent manner(P<0.05).β-eleostearic acid significantly promoted the release of LDH and promoted the apoptosis of JEG-3 cells(P<0.05),and the expression level of apoptosis related protein Caspase-3 and the expression ratio of Bax/Bcl-2 were up-regulated.It proved thatβ-eleostearic acid could induce the apoptosis of JEG-3 cells.In addition,β-eleostearic acid could increase the content of ROS in JEG-3 cells.The inhibitory effect ofβ-eleostearic acid on the relative activity of JEG-3 cells could be antagonized by ROS inhibitor NAC(P<0.05).Conclusion:β-eleostearic acid could induce the apoptosis of JEG-3 cells.Its effect was likely related to the production of ROS.