马鞭草总黄酮通过AKT/mTOR信号通路调控自噬抑制肝癌细胞增殖  

Total flavonoids of verbena officinalis L.regulate autophagy and inhibit hepatocellular carcinoma cell proliferation through AKT/mTOR pathway

作　　者：杨金旭[1] 孙自红[1] 徐琳[1] YANG Jinxu;SUN Zihong;XU Lin(Department of Nursing,Luohe Medical College,Henan Luohe 462002,China)

机构地区：[1]漯河医学高等专科学校护理系,河南漯河462002

出　　处：《现代肿瘤医学》2023年第14期2631-2638,共8页Journal of Modern Oncology

基　　金：河南省科技攻关计划资助项目(社会发展领域)(编号:162102310596)。

摘　　要：目的:分析马鞭草总黄酮(total flavonoids of verbena officinalis L.,TFV)对肝癌细胞自噬和增殖的影响,并探讨其机制。方法:MDC染色法检测自噬泡水平,Western印迹检测自噬相关蛋白水平,运用3-甲基腺嘌呤(3-MA)、雷帕霉素(RAPA)研究TFV对自噬、细胞增殖的影响,MTT法检测细胞存活率。裸鼠成瘤实验验证TFV对裸鼠体内瘤体的瘤重、瘤体积及自噬相关蛋白水平的影响。结果:TFV能增加HepG2、Huh-7细胞中MDC染色标记的荧光颗粒,并伴随浓度的上调明显增加,促进HepG2、Huh-7细胞中自噬蛋白Beclin-1、LC3 II/LC3 I水平表达,降低p-AKT/AKT、p-mTOR/mTOR水平(P<0.05)。与TFV组比较,TFV+3-MA组抑制率、LC3 II/LC3 I明显降低,p-mTOR/mTOR明显增加(P<0.05);与TFV组比较,TFV+RAPA组抑制率、LC3 II/LC3 I明显增加,p-mTOR/mTOR明显降低(P<0.05);与TFV组比较,TFV+3-MA组裸鼠瘤体LC3 II/LC3 I明显降低,瘤体质量、瘤体体积、p-mTOR/mTOR明显增加(P<0.05)。结论:TFV可通过抑制AKT/mTOR信号通路诱导细胞自噬来抑制肝癌细胞增殖。Objective:To analyze the effect of total flavonoids of verbena officinalis L.(TFV)on autophagy and proliferation of hepatocarcinoma cells,and to explore its mechanism.Methods:MDC staining method was used to detect the level of autophagy vesicles.Western blotting was used to detect autophagy-related protein levels.3-MA and Rapamycin(RAPA)were used to study the effects of TFV on autophagy and cell proliferation,and CCK-8 method was used to detect cell survival.The tumor formation experiment in nude mice verified the effects of TFV on tumor weight,tumor volume and autophagy-related protein levels in nude mice.Results:TFV could increase the fluorescent particles labeled by MDC staining in HepG2 and Huh-7 cells,and the fluorescent particles significantly increased with the increase of concentration.It could promote the expression of autophagy protein Beclin-1 and LC3 II/LC3 I in HepG2 and Huh-7 cells,and reduce the levels of p-AKT/AKT and p-mTOR/mTOR(P<0.05).Compared with the TFV group,the inhibition rate and LC3 II/LC3 I levels in the TFV+3-MA group were significantly reduced,and the p-mTOR/mTOR levels was significantly increased(P<0.05).Compared with the TFV group,the inhibition rate and LC3 II/LC3 I levels in the TFV+RAPA group were significantly increased,and the levels of p-mTOR/mTOR were significantly reduced(P<0.05).Compared with the TFV group,LC3 II/LC3 I levels of the tumor bodies of nude mice in the TFV+3-MA group decreased significantly,and tumor mass,tumor volume,and p-mTOR/mTOR increased significantly(P<0.05).Conclusion:TFV can inhibit the proliferation of hepatocellular carcinoma cells by inducing autophagy by inhibiting the AKT/mTOR signaling pathway.

关 键 词：马鞭草总黄酮 AKT/mTOR号通路 自噬 肝癌 细胞增殖 

分 类 号：R735.7[医药卫生—肿瘤]