熊果酸诱导人胃癌细胞株MGC-803发生凋亡和自噬  

Ursolic Acid Induces Apoptosis and Autophagy in Human Gastric Cancer Cell Line MGC-803

作　　者：陈伟妍 刘春英[1] CHEN Weiyan;LIU Chunying(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China)

机构地区：[1]辽宁中医药大学

出　　处：《中华中医药学刊》2019年第8期1832-1835,I0012,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81473569)

摘　　要：目的:观察熊果酸(UA)对胃癌细胞株MGC-803凋亡与自噬的影响。方法:体外培养人胃癌细胞株MGC-803,以CCK8法测定UA对MGC-803细胞活力的影响并筛选出UA最佳作用浓度和时间;采用倒置相差显微镜观察UA干预后MGC-803细胞的形态学改变;采用MDC染色法观察UA对MGC-803细胞自噬的影响;采用实时定量PCR法检测凋亡相关基因BAX、BCL2和自噬相关基因Beclin1、LC3Ⅱ的表达水平;采用WB法对上述凋亡及自噬相关基因进行蛋白水平的验证。结果:CCK8法显示UA能显著抑制MGC-803细胞增殖,并具有浓度和时间依赖性;倒置相差显微镜观察到UA干预后视野下MGC-803细胞数量显著减少,部分细胞的形态发生变化,呈现碎片化;MDC染色法显示UA干预后视野下MGC-803细胞的荧光信号显著增强;实时定量PCR法显示UA显著上调凋亡相关基因BAX水平以及自噬相关基因Beclin1、LC3Ⅱ水平(P<0.05),显著下调凋亡相关基因BCL2水平(P<0.05);WB法显示UA显著上调凋亡相关蛋白BAX水平以及自噬相关蛋白Beclin1、LC3Ⅱ水平(P<0.05),显著下调凋亡相关蛋白BCL2水平(P<0.05)。结论:熊果酸对人胃癌细胞株生长具有显著的抑制作用且呈剂量和时间依赖性,同时熊果酸诱导人胃癌细胞死亡过程中凋亡和自噬均被激活。Objective:To observe the effect of ursolic acid(UA) on apoptosis and autophagy in gastric cancer cell line MGC-803.Methods:Human gastric cancer cell line MGC-803 was cultured in vitro. The influence of UA on the viability of MGC-803 cells was determined by CCK8 method and the best concentration and time of UA were screened out. Inverted phase contrast microscope was used to observe the morphological changes of MGC-803 cells after UA intervention. MDC staining was used to observe the effect of UA on autophagy in MGC-803 cells. The expression levels of apoptosis-related genes BAX, BCL2 and autophagy-related genes Beclin1 and LC3Ⅱ were detected by real-time quantitative PCR. The apoptosis and autophagy related genes were verified by WB.Results:CCK8 showed that UA could significantly inhibit the proliferation of MGC-803 cells in a concentration and time dependent manner.The number of MGC-803 cells decreased significantly after UA intervention. Some cells undergo morphological changes, showing fragmentation.MDC staining showed that the fluorescence signal of MGC-803 cells increased significantly after UA intervention.Real-time quantitative PCR showed that UA significantly increased the levels of apoptosis-related genes BAX and autophagy-related genes Beclin1 and LC3Ⅱ(P<0.05), and significantly decreased the levels of apoptosis-related genes BCL2(P<0.05).WB assay showed that UA significantly increased the levels of apoptosis-related protein BAX and autophagy-related protein Beclin 1 and LC3Ⅱ(P<0.05), and significantly decreased the level of apoptosis-related protein BCL2(P<0.05).Conclusion:Ursolic acid has a significant inhibitory effect on the growth of human gastric cancer cell lines in a dose and time dependent manner. Ursolic acid induces apoptosis and autophagy activation in the process of human gastric cancer cell death.

关 键 词：熊果酸 人胃癌细胞株MGC-803 凋亡 自噬 

分 类 号：R285.5[医药卫生—中药学]