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右旋硫辛酸对人肝癌HepG2细胞生长增殖及其相关机制研究

刘晶, 黄严伟, 周波, 宋清清

刘晶, 黄严伟, 周波, 宋清清. 右旋硫辛酸对人肝癌HepG2细胞生长增殖及其相关机制研究[J]. 中国药科大学学报, 2017, 48(3): 348-354. DOI: 10.11665/j.issn.1000-5048.20170316
引用本文: 刘晶, 黄严伟, 周波, 宋清清. 右旋硫辛酸对人肝癌HepG2细胞生长增殖及其相关机制研究[J]. 中国药科大学学报, 2017, 48(3): 348-354. DOI: 10.11665/j.issn.1000-5048.20170316
LIU Jing, HUANG Yanwei, ZHOU Bo, SONG Qingqing. Effect of R-(+)-lipoic acid on growth, proliferation and related mechanism in human HepG2 cells[J]. Journal of China Pharmaceutical University, 2017, 48(3): 348-354. DOI: 10.11665/j.issn.1000-5048.20170316
Citation: LIU Jing, HUANG Yanwei, ZHOU Bo, SONG Qingqing. Effect of R-(+)-lipoic acid on growth, proliferation and related mechanism in human HepG2 cells[J]. Journal of China Pharmaceutical University, 2017, 48(3): 348-354. DOI: 10.11665/j.issn.1000-5048.20170316

右旋硫辛酸对人肝癌HepG2细胞生长增殖及其相关机制研究

Effect of R-(+)-lipoic acid on growth, proliferation and related mechanism in human HepG2 cells

  • 摘要: 研究天然抗氧化剂右旋硫辛酸对人肝癌HepG2细胞生长、增殖的影响及其相关机制。采用MTT检测细胞增殖情况,活性氧试剂盒分析细胞内ROS水平。流式细胞术和Hoechst 33258染色观察细胞凋亡和形态变化。Western blot检测凋亡、自噬以及相关通路蛋白表达,包括Bax、Bcl-2、caspase 3、PARP、ATG5、ATG7、LC3、Beclin1、mTOR、P70S6K、P38、P53、ERK、Akt、MEK等。结果表明,经不同浓度不同时间药物处理,右旋硫辛酸可抑制HepG2细胞增殖,提高细胞内ROS水平,并呈时间和剂量依赖性。右旋硫辛酸通过上调促凋亡蛋白Bax,激活caspase家族,从而激活凋亡效应蛋白caspase 3和PARP,同时右旋硫辛酸还可上调自噬相关蛋白ATG5、ATG7、Beclin1、LC3水平,抑制磷酸化mTOR和P70S6K,激活自噬。通路研究表明:右旋硫辛酸可上调磷酸化的P38和JNK促凋亡通路促进凋亡,抑制磷酸化Akt、ERK的表达。添加自噬抑制剂3-甲基腺嘌呤后,明显抑制自噬发生。因此,右旋硫辛酸可能通过调控P38/AMPK-JNK,PI3K/Akt和Ras/Raf/MEK/ERK途径激活自噬,诱导细胞凋亡。
    Abstract: To study the effect of antioxidants R-(+)-lipoic acid(R-LA)on cells growth, proliferation and related mechanisms in human HepG2 cells lines. MTT was used to measure cells growth and proliferation. Reactive oxygen species(ROS)kit was used to analyze ROS level. Cell apoptosis and cell morphological changes were observed by flow cytometry and Hoechst 33258 test. Protein expression levels of apoptosis, autophagy and related pathway were analyzed through Western blot, including Bax, Bcl-2, caspase 3, PARP, ATG5, ATG7, LC3, Beclin1, mTOR, P70S6K, P38, P53, ERK and Akt etc. Results showed that cell growth and proliferation were inhibited in a dose- and time-dependent manner after being treated by lipoic acid. R-LA could increase ROS production, pro-apoptosis proteins Bax levels, activated caspase family and PARP. Meanwhile, R-LA could up-regulate the levels of autophagy-related proteins including ATG5, ATG7, Beclin1 and LC3, and down-regulate phospho-mTOR and P70S6K levels. Signal pathway results showed that R-LA could up-regulate phospho-P38 and phospho-JNK levels, and decrease phospho-Akt and phospho-ERK. When adding 3-methyladenine, autophagy was inhibited. Thus, R-LA might activate autophagy and induce apoptosis by P38/AMPK-JNK, PI3K/AKT and Ras/Raf/MEK/ERK pathways.
  • [1] Flavia N,Mike FQ,Cristina S.Lipoic acid:a unique antioxidant in the detoxification of activated oxygen species[J].Plant Physiol Bioch,2002,40(25):463-470.
    [2] Zhang JY,Ji SZ.The research progress of lipoic acid[J].Mod Chem Ind(现代化工),2012,32(5):32-33.
    [3] Al-Rasheed NM,Al-Rasheed NM,Attia HA,et al.Adverse cardiac responses to alpha-lipoic acid in a rat-diabetic model:possible mechanisms[J]?Physiol Biochem,2013,69(4):761-78.
    [4] Kim JI,Cho SR,Lee CM,et al.Induction of ER stress-mediated apoptosis by α-lipoic acid in A549 cell lines Korean[J].Thorac Cardiov Surg,2012,45(1):1-10.
    [5] Simbula G,Columbano A,Ledda-Columbano GM,et al.Increased ROS generation and p53 activation in α-lipoic acid-induced apoptosis of hepatoma cells[J].Apoptosis,2007,12(1):113-123.
    [6] Feng QF,Bi L,Yan XJ,et al.Inhibition of tetramethypyrazine on proliferation of HepG2 cells and its effects on the pathway of mitochondrial apoptosis[J].J China Pharm Univ(中国药科大学学报),2015,46(3):350-354.
    [7] Krueger A,Baumann S,Peter H.Flice inhibitory proteins:regulators of death receptor mediated apoptosis[J].Mol Cell Biol,2001,24(21):8247-8254.
    [8] Park JY,Song JY,Kim HM.et al.p53-Independent expression of wildtype p53-induced phosphatase 1(Wip1)in methylmethane sulfonate-treated cancer cell lines and human tumors[J].Cell Biol,2012,44(5):896-904.
    [9] Schwabe RF,Brenner,DA.Mechanisms of liver injury.I.TNF-alpha-induced liver injury:role of IKK,JNK,and ROS pathways[J].AM J Physiol-Gastr L,2006,290(4):583-589.
    [10] Boldt S,Weidleand UH,W Kolch et al.The role of MAPK pathways in the action of chemotherapeutic drugs[J].Carcinogenesis,2002,23(11):1831-1838.
    [11] Zarubin T, Han J. Activation and signaling of the p38 MAP kinase pathway[J] Cell Res,2005,15:11-18.
    [12] Park GB,Choi Y,Kim YS,et al.ROS-mediated JNK/p38-MAPK activation regulates Bax translocation in sorafenib-induced apoptosis of EBV-transformed B cells[J].Int J Oncol,2014,44(3):977-985.
    [13] Park GB,Kim YS,Lee HK,et al.Reactive oxygen species and p38 MAPK regulate Bax translocation and calcium redistribution in salubrinal-induced apoptosis of EBV-transformed B cells[J].Cancer Lett,2011,313(2):235-248.
    [14] Yan TT,Zhao Y,Zhang X,et al.Astaxanthin inhibits acetaldehyde-induced cytotoxicity in SH-SY5Y cells by modulating Akt/CREB and p38MAPK/ERK signaling pathways[J].Mar Drugs,2016,14(3):56.
    [15] Dong YJ,Gao WJ.Bcl-2,Bax,caspase 3 in the role of apoptosis and their relationships[J].Chin J Gerontol(中国老年医学杂志),2012,32(21):4828-4830.
    [16] kafara P,lcard P,Guillamin M.Lipoic acid decreases Mcl-1,Bcl-xL and up regulates Bim on ovarian carcinoma cells leading to cell death[J].J Ovarian Res,2015,8(10):36.
    [17] Koike N,Takeyoshi I,Ohki S,et al.Effects of adding P38 mitogen-activated protein-kinase inhibitor to celsior solution in canine heart transplantation from non-heart-beating donors[J].Transplantation,2004,77(2):286-292.
    [18] Hashimoto N,Takeyoshi I,Tsutsumi H,et al.Effects of a bradykinin B(2)receptor antagonist on ischemia-reperfusion injury in a canine lung transplantation model[J].J Heart Lung Transplant,2004,23(5):606-613.
    [19] Camera E,Mastrofrancesco A,Fabbri C,et al.Astaxanthin,canthaxanthin and beta-carotene differently affect UVA-induced oxidative damage and expression of oxidative stress-responsive enzymes[J].Exp Dermatol,2009,18(3):222-231.
    [20] Lum JJ, Bauer DE, Kong M, et al. Growth factor regulation of autophagy and cell survival in the absence of apoptosis[J].Cell,2005,120(2):237-248.
    [21] Zhao F,Huang WW,Zhang Z,et al.Triptolide induces protective autophagy through activation of the CaMKKβ-AMPK signaling pathway in prostate cancer cells[J].Oncotarget,2016,7(5):5366-5382.
    [22] laplante M,Sabatini DM.mTOR signaling in growth control and disease[J].Cell,2012,149(2):274-293.
    [23] Jung CH,Ro SH,Cao J,et al.mTOR regulation of autophagy[J].FEBS,2010,584(7):1287-1295.
    [24] Scherz-Shouval R,Elazar Z.Regulation of autophagy by ROS:physiology and pathology[J].Trends Brioche Sci,2011,36(1):8-30.
    [25] He CC,Klionsky DJ.Regulation mechanisms and signaling pathways of autophagy[J].Annu Rev Genet,2009,43(9):67-93.
    [26] Inoki K,Li Y,Zhu T,et al.TSC2 is phosphorylated and inhibited by Akt and suppresses mTOR signaling[J].Nat Cell Biol,2002,4(9):648-657.
    [27] Ma L,Chen Z,Erdjument-Bromage H,et al.Phosphorylation and functional inactivation of TSC2 by Erk:implications for tuberous sclerosis and cancer pathogenesis[J].Cell,2005,121(2):179-193.
    [28] Peterson TR,Laplante M,Thoreen CC,et al.DEPTOR is an mTOR inhibitor frequently overexpressed in multiple myeloma cells and required for their survival[J].Cell,2009,137(5):673-886.
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出版历程
  • 刊出日期:  2017-06-24

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