藤黄酸对食管鳞状细胞癌TE-1细胞caspase-3活性和细胞自噬作用影响的研究  

Study on the effect of gambogic acid on caspase-3 activity and autophagy of esophageal squamous-cell carcinoma TE-1 cells

作　　者：刘文粤 黄淼龙[1] 万仁平[1] 陈翀[1] 陈慧勇[1] LIU Wenyue;HUANG Miaolong;WAN Renping;CHEN Chong;CHEN Huiyong(Department of Thoracic Surgery,YueBei People’s Hospital,Shaoguan 512026,China;不详)

机构地区：[1]粤北人民医院,广东韶关512026

出　　处：《现代医院》2020年第7期1048-1052,共5页Modern Hospitals

基　　金：韶关市科技计划项目(2018sn106)。

摘　　要：目的本研究选取食管鳞状细胞癌细胞株为研究对象,探讨藤黄酸对食管鳞状细胞癌TE-1细胞caspase-3活性和细胞自噬作用影响的研究,旨在明确藤黄酸的抗癌作用及机制。方法将食管鳞状细胞癌TE-1细胞随机分为6组,每组细胞分别加入0μg/mL、4μg/mL、6μg/mL、8μg/mL、10μg/mL各浓度的藤黄酸提取液各10μL,同时还设立一组为10μg/mL藤黄酸+caspase抑制剂z-VAD联合作用组,每组设4个复孔;用caspase-3活性检测试剂盒对caspase-3活性的活性进行检测,运用CCK-8试剂盒对藤黄酸各浓度处理组以及藤黄酸+z-VAD组细胞的活性进行检测。运用细胞自噬实验对藤黄酸各浓度处理组(0μg/mL、4μg/mL、6μg/mL、8μg/mL、10μg/mL)中的细胞自噬情况进行检测。结果caspase-3活性检测实验结果显示,随着藤黄酸浓度的上升,caspase-3活性随之升高,当藤黄素浓度为10μg/mL时caspase-3活性到达顶峰,而当加入caspase-3抑制剂z-VAD后,caspase-3活性受到了明显抑制。CCK-8试剂盒检测结果表明,随着藤黄酸浓度的增加,TE-1细胞的活性抑制率逐渐升高,当用藤黄酸浓度为10μg/mL时对食管鳞状细胞癌TE-1细胞活性的抑制率最高,而当加入caspase-3抑制剂z-VAD后,TE-1细胞的活性的抑制率显著降低。细胞自噬实验结果表明,各藤黄酸处理组未见显著差异。结论藤黄酸能够诱导caspase-3活性促进TE-1细胞凋亡,且呈剂量依赖性,但对细胞的自噬作用无显著影响。Objective To investigate the effect of gambogic acid on caspase-3 activity and autophagy of Esophageal squamous-cell Carcinoma TE-1 cells,so as to clarify GA anticancer effect and mechanism.Methods Esophageal squamous cell carcinoma TE-1 cells were randomly divided into 6 groups,each group of cells was added 10μL of 0μg/mL,4μg/mL,6μg/mL,8μg/mL,10μg/mL,and we also set up a group of 10μg/mL GA+caspase inhibitor z-VAD combination group,each concentration of GA extract had four duplications.We used caspase-3 activity assay kit to detected the active of caspase-3 activity,and used CCK-8 cell viability assay kit to detect the cell viability.Results Caspase-3 activity assay results showed that the caspase-3 activity were significantly increased with increasing concentrations of GA,when GA concentration was 10μg/mL the caspase-3 activity was the most.When added to caspase-3 inhibitor z-VAD,caspase-3 activity has been inhibited.CCK-8 kit test results showed that with increasing concentrations of GA,TE-1 cell activity decreased,when using GA concentration of 10μg/mL of esophageal squamous cell carcinoma TE-1 cell activity the highest inhibition rate,and when added to caspase-3 inhibitor z-VAD,activity inhibition rate TE-1 cells was significantly reduced.Autophagic cell experiments showed that there was no significant difference in the GA between treatment groups.Conclusion GA could induce TE-1 cells apoptosis in a dose-dependent manner via increasing the caspase-3 activity,but had no significant effect on autophagy.

关 键 词：食管鳞状细胞癌 藤黄酸 CASPASE-3 细胞凋亡 

分 类 号：R735.1[医药卫生—肿瘤]