熊果酸对肺癌细胞A549自噬相关蛋白的影响  

Effect of Ursolic Acid on Autophagy-Related Proteins in Lung Cancer Cell Line A549

作　　者：陈姝瑾 王淳[2] 刘春英[2] CHEN Shujin;WANG Chun;LIU Chunying(Graduate School of Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China;College of Integrated Traditional Chinese and Western Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,Liaoning,China)

机构地区：[1]辽宁中医药大学研究生学院,辽宁沈阳110847 [2]辽宁中医药大学中西医结合学院,辽宁沈阳110847

出　　处：《中华中医药学刊》2020年第6期86-90,I0020,共6页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金(81973735,81774184)。

摘　　要：目的研究自噬在熊果酸抑制人肺癌A549细胞增殖中的作用及机制。方法应用CCK8法检测熊果酸对A549细胞增殖的影响;实时荧光定量PCR法检测自噬相关基因LC3、Beclin-1及p62的表达情况;Western Blot检测自噬相关蛋白LC3、Beclin-1及p62的表达情况。采用自噬抑制剂3-甲基腺嘌(3-MA)观察熊果酸对A549细胞增殖的抑制作用。结果熊果酸可以显著抑制人肺癌A549细胞的活力(P<0.01)且随着熊果酸剂量增加,对细胞的生长抑制率显著上升。熊果酸可诱导A549细胞自噬发生,诱导自噬相关基因、蛋白LC3Ⅱ/Ⅰ、Beclin-1表达的增加;p62表达降低(P<0.01);3MA使得自噬相关基因、蛋白LC3Ⅱ/Ⅰ、Beclin-1表达明显降低;p62表达明显增加(P<0.01)。自噬抑制剂3-MA提高了熊果酸对A549细胞的抑制作用(P<0.01)。结论熊果酸抑制A549细胞的增殖,可能通过调节LC3Ⅱ/Ⅰ、Beclin-1及p62蛋白表达诱导细胞发生自噬。自噬抑制剂3-MA能够增强熊果酸抑制A549细胞的增殖抑制作用,有望为肺癌的临床治疗提供新的联合治疗方案。Objective To study the role and mechanism of autophagy in the inhibition of ursolic acid on the proliferation of human lung cancer A549 cells.Methods The effect of ursolic acid on the proliferation of A549 cells was detected by CCK8 method.The expressions of autophagy-related genes LC3,Beclin-1 and p62 were detected by real-time fluorescent quantitative PCR.The expressions of the autophagy-related proteins LC3,Beclin-1 and p62 were detected by Western blot.The inhibitory effect of ursolic acid on the proliferation of A549 cells was observed by the autophagy inhibitor 3-methyladenine(3-MA).Results Ursolic acid significantly inhibited the viability of human lung cancer A549 cells(P<0.01).With the increase of ursolic acid dose,the inhibition rate of cell growth was significantly increased.Ursolic acid induced autophagy in A549 cells,induced the increase of expressions of autophagy-related genes,protein LC3Ⅱ/Ⅰand Beclin-1 as well asthe decrease of p62 expression(P<0.01).3-MA significantly decreased the expressions of autophagy-related genes,protein LC3Ⅱ/Ⅰand Beclin-1 and increased p62 expression significantly(P<0.01).The autophagy inhibitor 3-MA increased the inhibitory effect of ursolic acid on A549 cells(P<0.01).Conclusion Ursolic acid inhibits the proliferation of A549 cells,which may induce autophagy by regulating the expressions ofLC3Ⅱ/Ⅰ,Beclin-1 and p62 protein.The autophagy inhibitor 3-MA can enhance the inhibitory effect of ursolic acid on the proliferation of A549 cells,and it is expected to provide a new combination therapy for the clinical treatment of lung cancer.

关 键 词：熊果酸 细胞增殖抑制 BECLIN-1 P62 自噬 

分 类 号：R285.5[医药卫生—中药学]