迷迭香酸甲酯通过抑制Akt/mTOR信号通路诱导人肝癌细胞凋亡  

Methyl rosmarinate induces cell apoptosis in human hepatocellular carcinoma cells via inhibiting the Akt/mTOR signaling pathways

作　　者：鲜瑶[1] 江伟 刘润坤 涂康生[3] 高士杰 王军[4] 张雷[5] XIAN Yao;JIANG Wei;LIU Runkun;TU Kangsheng;GAO Shijie;WANG Jun;ZHANG Lei(Department of Nutrition,The First Affliliated Hospital of Xi’an Jiaotong University,Xi’an 710061;School of Nursing,Shaoyang University,Shaoyang 422000;Department of Hepatobiliary Surgery,The First Affliliated Hospital of Xi’an Jiaotong University,Xi’an 710061;Department of Emergency and Intensive Care Unit(East District),The First Affliliated Hospital of Xi’an Jiaotong University,Xi’an 710089;Department of Geriatric Surgery,The First Affliliated Hospital of Xi’an Jiaotong University,Xi’an 710061,China)

机构地区：[1]西安交通大学第一附属医院营养科,陕西西安710061 [2]邵阳学院护理学院,湖南邵阳422000 [3]西安交通大学第一附属医院肝胆外科,陕西西安710061 [4]西安交通大学第一附属医院急诊与重症医学科(东院区),陕西西安710089 [5]西安交通大学第一附属医院老年外科,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2023年第5期802-808,共7页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：陕西省自然科学基础研究计划(No.2023-JC-QN-0915);陕西省重点研发计划(No.2020SF-061,No.2020KW-054);西安交通大学第一附属医院院基金(No.2021ZYTS-37,No.2019ZYTS-07)资助。

摘　　要：目的探讨迷迭香酸甲酯(methyl rosmarinate,MR)诱导人肝癌细胞(HCC)Hep-3B、SK-Hep1凋亡的作用及其机制。方法CCK-8法检测各浓度MR(0~200μmol/L)对Hep-3B、SK-Hep1、MIHA细胞的增殖-毒性作用;光学显微镜观察不同浓度MR处理三种细胞后的形态变化;EdU实验及流式细胞术分别检测Hep-3B、SK-Hep1细胞增殖及凋亡情况;Transwell迁移和侵袭实验检测MR对Hep-3B、SK-Hep1细胞迁移与侵袭能力的影响;Western blotting检测凋亡、上皮间质转化(epithelial-mesenchymal transition,EMT)及Akt/mTOR信号通路相关蛋白的表达情况。结果不同浓度MR(0~200μmol/L)处理Hep-3B、SK-Hep1细胞48 h,其细胞活性呈浓度依赖性显著降低(P<0.01),而对MIHA细胞活性无显著影响(P>0.05),Hep-3B、SK-Hep1细胞MR的IC 50分别为102.5μmol/L和99.3μmol/L。MR(0~150μmol/L)处理后显著抑制Hep-3B、SK-Hep1细胞增殖(P<0.05),通过光学显微镜观察到Hep-3B、SK-Hep1贴壁细胞死亡脱落和细胞外形凹陷萎缩,而MIHA细胞形态无明显改变。与对照组相比,100、150μmol/L MR诱导Hep-3B、SK-Hep1细胞凋亡,凋亡蛋白Bax和cleaved PARP的表达显著增加(P<0.05),而抗凋亡蛋白Bcl-2的表达显著下降(P<0.05);且HCC细胞的迁移和侵袭均被抑制,E-cadherin的表达显著增加,N-cadherin和Vimentin的表达降低(P<0.05)。Western blotting检测结果显示,经MR处理的Hep-3B和SK-Hep1的p-Akt和p-mTOR的表达显著降低且呈剂量依赖性,这提示MR可能通过抑制Akt/mTOR信号通路发挥抗癌作用。结论MR促进Hep-3B、SK-Hep1细胞凋亡,其作用机制可能与抑制Akt/mTOR信号通路有关。Objective To investigate the cell death-inducing effect of methyl rosmarinate(MR)on human hepatoma Hep-3B and SK-Hep1 cells and their potential mechanisms.Methods The effects of MR on the viability of Hep-3B,SK-Hep1 and MIHA cells were determined by cell counting kit-8(CCK-8)assay.The morphological changes of three kinds of cells treated with different concentrations of MR were observed by optical microscopy.EdU assay and flow cytometry were used to detect the proliferation and apoptosis of Hep-3B and SK-Hep1 cells.Transwell assay was used to study the effects of MR on the migration and invasion of Hep-3B and SK-Hep1 cells.Western blotting was used to evaluate the protein expression levels of apoptosis,EMT and Akt/mTOR signaling pathways.Results After treated with different concentrations of MR(0~200μmol/L)for 48 h,Hep-3B and SK-Hep1 cells activities were significantly decreased in a concentration-dependent manner(P<0.01),while there was no significant effect on MIHA cell activity(P>0.05),and the IC 50 of Hep-3B and SK-Hep1 cells were 102.5 and 99.3μmol/L,respectively.MR treatment(0-150μmol/L)significantly inhibited the proliferation of Hep-3B and SK-Hep1 cells(P<0.05),while cell detachment and shrinkage were observed by optical microscopy on the Hep-3B and SK-Hep1 cells,while the morphology of MIHA cells was not changed.Compared with the control group,MR(100,150μmol/L)induced apoptosis in Hep-3B and SK-Hep1 cells,the expression levels of the pro-apoptotic proteins Bax and cleaved PARP were significantly increased(P<0.05),while the expression level of the anti-apoptotic protein Bcl-2 was significantly decreased(P<0.05).MR(100,150μmol/L)also inhibited the migration and invasion of HCC cells,significantly increased the expression of E-cadherin and decreased the expression of N-cadherin and Vimentin compared with the control group(P<0.05).Finally,Western blotting results showed that the expressions of p-Akt and p-mTOR in Hep-3B and SK-Hep1 treated by MR were significantly reduced in a dose-dependent manner,sugg

关 键 词：迷迭香酸甲酯 原发性肝癌 增殖 迁移 凋亡 Akt/mTOR通路 

分 类 号：R735.7[医药卫生—肿瘤]