摘要
【目的】“五行汤”由牛蒡、白萝卜、胡萝卜、干香菇和干白萝卜叶配制而成,民间服用有一定的抗肿瘤效果,值得深入研究。在大力发展传统医药的今天,本课题结合现代分子生物学技术观察研究五行汤的抗癌效果及其机制。
【方法】1、不同批次五行汤稳定性鉴定:紫外-可见光吸收光谱分析和成分定性检测,比较不同产地和不同贮存时间的五行汤组分相似度;不同批次五行汤处理肿瘤细胞,设立浓度梯度,MTT法检测比较抑瘤作用。2、五行汤促肿瘤细胞SGC-7901死亡方式检测:荧光染色和电镜观察细胞形态学变化;AnnexinV-FITC/PI双标法检测凋亡率。3、五行汤促SGC-7901细胞凋亡分子机制的探究:蛋白印迹检测凋亡途径蛋白变化;钙离子特异性荧光探针检测胞浆钙离子浓度变化;吖啶橙染色检测溶酶体膜完整性;Annexin V结合实验和CCK-8法检测不同抑制剂对细胞凋亡和存活的影响。
【结果】选取不同产地原料,在不同时间按标准流程配制不同批次五行汤;紫外-可见光吸收光谱显示不同批次五行汤吸收峰波形相似。不同批次五行汤主要成分定性实验结果一致:蛋白质、酚类、黄酮检测均为阳性、生物碱检测均为阴性。不同批次五行汤均可抑制肿瘤细胞SGC-7901生长,且抑制作用接近。PI荧光染色和电镜观察到五行汤处理的SGC-7901细胞呈凋亡特征;AnnexinV-FITC/PI双标流式检测到五行汤促SGC-7901细胞凋亡作用。免疫印迹检测半胱天冬氨酸酶(caspase)家族蛋白无活化迹象;Annexin V结合实验检测到广谱caspase抑制剂对细胞凋亡没有影响。荧光探针检测到胞浆钙离子浓度升高;胞内钙离子拮抗剂Bapta AM可以明显降低五行汤诱导的SGC-7901细胞凋亡(P＜0.05),且对细胞保护作用呈浓度依赖性。吖啶橙(AO)染色观察到溶酶体膜渗漏;胞浆蛋白免疫印迹检测到溶酶体释放蛋白酶cathepsin D和cathepsin B至胞浆;天冬氨酸蛋白酶cathepsin D抑制剂对五行汤诱导的细胞凋亡没有影响;半胱氨酸蛋白酶cathepsin B、L、S抑制剂可以明显减少细胞凋亡(P＜0.05),且对细胞的保护作用呈浓度依赖性。胞内钙离子拮抗剂Bapta AM可以减少溶酶体膜渗漏。JC-1染色检测出线粒体膜电位降低;胞浆蛋白免疫印迹发现线粒体没有释放细胞色素C(cyto C)而选择性释放凋亡诱导因子(AIF);核蛋白免疫印迹显示AIF没有入核;AIF抑制剂和钙蛋白酶calpain抑制剂均对五行汤诱导的SGC-7901细胞凋亡没有保护作用。
【结论】一定范围内的不同批次五行汤组分相似度较高,抑瘤活性相对稳定;同时五行汤可以通过促肿瘤细胞SGC-7901凋亡发挥抑瘤作用;对其机制研究显示胞浆钙离子浓度升高和溶酶体相关凋亡蛋白是该凋亡过程中关键因素,提示五行汤经钙离子/溶酶体介导的caspase非依赖凋亡通路诱导细胞凋亡。
Objective: Wuxing soup is made of burdock, white radish, carrot, dry mushroom and the leaves of white radish, which has some antitumor effect in folk medicine. On the background of promoting the development of Chinese traditional medicine energetically, this study based on the modern molecular biology techniques aims to explore the anti-cancer effect and mechanisms of Wuxing soup.
Methods: First, investigation on the stability of Wuxing soup in different groups: the similarity calculation was analysed by UV-VIS spectroscopy and qualitative test; the anti-cancer effect was dectected by MTT assay. Second, detection of death style: morphological change of SGC-7901 was observed by fluorescence staining and transmission electron microscopy. Measurement on the apoptotic rate was detected by FCM. Third, study on the mechanisms of Wuxing soup-induced apoptosis: the key proteins in apoptotic pathway were tested by Western Blot; the concentration of calcium was measured by fluorescence probe; the integrity of lysosome membrane was inspected by Acridine Orange staining; the effect of inhibitors on cell apoptotic rate and survival rate were analysed by Annexin V-binding ELISA and CCK-8 respectively.
Results: 1) Optical absorption shape of different batches of Wuxing soup was similar dectected by UV-VIS spectrophotometry. 2) Components in different batches of Wuxing soup were approximate the same by qualitative detection. 3) The anti-cancer effect of different batches of Wuxing soup was similar. 4) Apoptosis features of SGC-7901 under Wuxing soup treatment were observed by transmission electron microscopy and PI fluorescence staining; and apoptosis was confirmed by Annexin V-FITC/PI staining dectected by FCM. 5) Caspases had no sign of activation and pan-caspase inhibitor had no effect on the apoptosis of SGC-7901. 6) The concentration of calcium in the cytosol was elevated and calcium inhibitor Bapta AM could reduce the apoptotic rate and protect SGC-7901 cells in a dose dependent manner. 7) The leakege of lysosome membrane was observed by AO staining and the lysosome associated apoptotic protein cathepsin D and cathepsin B were released in cytosol. 8) The inhibitor of cathepsin B, L, S exerted protection in the survival of SGC-7901 under Wuxing soup treatment in a dose dependent manner. 9) The cell mitochondrial transmembrane potential was decreased and mitochondria selectively released AIF. 10) AIF had no sign of translocation to the nucleus. 11) The inhibitors of AIF, calpain and cathepsin D exerted no protection on the apoptosis induced by Wuxing soup.
Conclusion: The components of Wuxing soup in different batches had a high similarity within a certain range. Wuxing soup had an effective inhibition on cancer cells, and could induce SGC-7901 cell apoptosis through calcium mediated and lysosome involved caspase-independent pathway.
引文
[1]赖春丽,黄婉锋,祝晨.等.抗肿瘤中药新药开发探讨[J].中药材.2003,26(9):677-680
[2]单厚昌,马伟光,高中祖.我国天然抗肿瘤药物的研究进展[J].现代中西医结合杂志.2005,14(6):825-827
[3]刘斌.和黄医药携手德国默克发力植物药自主研发[J].中国医药导报.2007,4(1):22-23
[4]何振伟.抗肿瘤植物药的应用进展[J].医药导报.2006,25(12):1303-1305
[5]陈德轩,潘立群.抗肿瘤植物蓟藤梨根[J].中药材.2004,27,(2):86-87
[6]刘耕陶.中药药理研究与药物创新[M].中国协和医科大学出版社2006:7
[7]李大鹏.康莱特注射液药效学及临床研究进展[J].中国新药与临床杂志.2007,26(10):778-782
[8]李仪奎 中药药理实验方法学[M].上海:上海科学技术出版社2006:22
[9]刘建勋,丛伟红.我国植物药(中药)研究的问题与建议[J].中国中药杂志.2002,27(7):484-486
[10]周岱翰.中医肿瘤学临床研究现状与评析[J].新中医.2006,38(6):4-6
[11]高宣亮,秦洁贞.食物的药理和疗效[M].北京:中国医药科技出版社2003
[12]莫非,胡晶莹,甘愉,等.五行汤对小鼠黑色素瘤抑瘤作用和免疫功能的影响[J].中国癌症杂志,2008,18(3):186-189
[13]莫非,胡晶莹,甘愉,等.五行汤抗肿瘤及经钙离子介导的促肿瘤细胞凋亡作用[J].中药材,已接收
[14]季宇彬,高世勇,孔琪,等.海嘧啶对SGC-7901人胃癌细胞凋亡的影响[J].中草药.2001,32(10):901-904
[15]周林,艾中立,刘志苏,等.复方苦参水溶液体外对人结肠癌SW480的影响[J].癌症.2000,19(4):337
[16]齐丽莎,孙保存.细胞自噬及其在肿瘤发生过程中的作用[J].国际肿瘤学杂志.2006,33(12):906-908
[17]Herman AA,Johnson DE,Singh SV,etal.Suforaphane causes autophagy to inhibit release of cytoehrome C and apoptosis in human prostate cancer cell[J].Cancer Res.2006,66(11):5828-5835
[18]Okada H,Mak TW.Pathway of apoptotic and non apoptotic cell death in tumor cells[J].Nat Rev Cancer.2004,4:592-603
[19]Assuncao GC,Linden R.Programmed cell death:Apoptosis and alternative deathstyles[J].Eur.J.Biochem.2004,271:1638-1650
[20]尤红,王宝恩,王泰龄.复方对肝星状细胞的增殖利凋亡的干预作用[J].中华肝脏病杂志。2000,8(2):78-80
[21]堵年生,王晓梅,王新玲,等.天然药物化学实验及学习指导[M].北京:科学出版社.2006
[22]彭心昭,陈英,朴英杰.自噬的抑制影响长春新碱诱导的肝癌细胞自噬性凋亡[J].肿瘤.2004,24(1):32-35
[23]Jun DY,Kim JS,Park HS,et al.Apoptogenic activity of auraptene of Zanthoxylum schinifolium toward human acute leukemia Jurkat T cells is associated with ER stress-mediated caspase-8 activation that stimulates mitochondria-dependent or -independent caspase cascade[J].Carcinogenesis.2007,(28) 6:1303-1313
[24]郑晓亮,刘雪莉,钱伯初.植物提取物诱导肿瘤细胞凋亡机制研究进展[J].中国药理学通报.2004,20(11):1201-5
[25]邵世光,赵忠顺.牛蒡根的药用研究进展[J].中医药学报.2006,34:24-25
[26]田甜.肿瘤细胞凋亡及植物药活性成分的干预效应研究进展[J].湖南中医学院学报.2005,25(4):62-65
[27]张俊文,王丕龙,汤为学.大豆异黄酮诱导人胃癌细胞株SGC-7901凋亡的研究[J].重庆医科大学学报.2004,29(3):329-331
[28]徐炎,蔡宇,梁少玲,等作用于细胞信号传导通路的抗肿瘤中药研究进展[J].上海中医药杂志2005,39(8):63-65
[29]苏勉诚,戴梅,李晓英,等中药复方癌宁对人胃腺癌细胞凋亡基因表达的影响[J].中药材,2002,(25):563-565
[30]Chan AS,Yip EC,Yung LY,et al.Immuno-regulatory Effects of CKBM on the Activities of Mitogen-Activated Protein Kinases and the Release of Cytokines in THP-1 Monocytic Cells[J].Biol.Pharm.Bull.2005,28(9):1645-1650
[31]Dong S,Inoue A,Zhu Y,et al.Activation of rapid signaling pathways and the subsequent transcriptional regulation for the proliferation of breast cancer MCF-7 cells by the treatment with an extract of Glycyrrhiza glabra root[J].Food Chem Toxicol.2007,(45):2470-2478
[32]Stander BA.,Marais S,Steynberg TJ,et al.Influence of Sutherlandia frutescens extracts on cell numbers,morphology and gene expression in MCF-7 cells[J].J Ethnopharmacol.2007,(112):312-318
[33]Lin YL,Lee TF,Huang YJ,Huang YT,et al.Inhibitory effects of Ligusticum chuanxiong on the proliferation of rat hepatic stellate cells[J].J Gastroenterol Hepatol.2006,(21):1257-1265
[34]Steinkamp-Fenske K,Bollinger L,Voller N,et al.Ursolic acid from the Chinese herb Danshen(Salvia miltiorrhiza L.) upregulates eNOS and downregulates Nox4 expression in human endothelial cells[J].Atherosclerosis.2007,(195):104-111
[35]叶人,蔡慎初,尹丽慧,等.扶正消瘕汤治疗胃癌的机制研究[J].中国中药杂志.2007, (32):1602-1604
[36]Shigeomi S,Toku K,Noboru M.Role of Bcl-2 family proteins in a non-apoptotic programmed cell death dependent on autophagy genes[J].Nat Cell Biol.2004,6:1221-1228
[37]Maiuri MC,Zalckvar E,Kimchi A,et al.Self-eating and self-killing:crosstalk between autophagy and apoptosis[J].Nature.2007,8:741-753
[38]李晓波,季莉莉,王钊.中药对细胞凋亡影响的研究进展[J].中药材.2002,25(2):135-139
[39]李勇,凌昌全.中药对肿瘤发生过程中信号转导异常的调整作用[J].中国中西医结合杂志.2003,23(11):876-879
[40]谢广茹,吴雄志.中药诱导肝癌细胞凋亡的研究进展[J].临床肿瘤学杂志。2007,12(3):225-227
[41]Green DR.Apoptotic pathways:paper wraps stone blunts scissors[J].Cell.2000,102(1):1-4.
[42]Nakagaws T,Zhu H,Morishima N,et al.Caspase-12 mediated endoplasmic-reticulum-specific apoptosis and cytotoxicity by a myloid -P[J].Nature.2000,403:98-103
[43]McConkey D J,Hartzell P,Nicotera P,et al.Stimulation of endogenous endonuclease activity in hepatocytas exposed to oxidative stress[J].Toxicol Lett,1988,42(2):123-130
[44]McConkey D J,Hartzell P,Amador-Perez JF,et al.Calcium-dependent killing of immature thymocytes by stimulation via the CD-3/T cell receptor complex[J].J Immunol.1989,43:1801-1806
[45]McConkey DJ,Aguilar SM,Hartzell P,et al.Induction of DNA fragmentation in chronic B-lymphocytic leukemia cells[J].J Immunol.1991,146(3):1072-1076
[46]Aw TY,Nicotera P,Manzo L,et al.Tributyltin stimulates apoptosis in rat thymocytes[J].Arch Biochem Biophys.1990,283(1):46-50
[47]Perotti M,Toddei F,Mirabelli F,et al.Calcium-dependent DNA flagmentation in human synovial cells exposed to cold shock[J].FEBS Lett.1990,259(2):331-334
[48]McConkey D J,Lin Y,Nutt LK,etal.Cardiac glycosides stimulate Ca increases and apoptosis in androgen-independent,metastatic human prostate adenocarcinoma cells[J].Cancer Res,2000,60(14):3807-3812
[49]Zhivotovsky B,Nicotera P,Bellomo G,etal.Ca~(2+) and endonuclease activation in radiation-induced lymphoid cell death[J].Exp Cell Res,1993,207(1):163-170
[50]Zhivotovsky B,Cedervall B,Jiang S,etal.Involvement of Ca~(2+) in the formation of high molecular weight DNA fragments in thymocyte apoptosis[J].Biochem Biophys Res Commun.1994,202(1):120-127
[51]Jiang S,Chow SC,McCabe MJ,etal.Orrenius S Lack of Ca~(2+) involvement in thymocyte apoptosis induced by chelation of intracellular Zn~(2+)[J].Lab Invest.1995,73(1):111 - 117
[52]Brunk UT,Neuzil J,Eaton JW.Lysosomal involvement in apoptosis[J].Redox Report.2001,6(2):91-97.
[53]Cirman T,Oresié K,Mazovec GD,et al.Selective disruption of lysosomes in HeLa cells triggers apoptosis mediated by cleavage of Bid by multiple papain-like lysosomal cathepsins[J].J Biol Chem.2004,279(5):3578-3587.
[54]Guicciardi ME,Leist M,Gores GJ.Lysosomes in cell death[J].Oncogene.2004,23(16):2881-2890.
[55]Michallet MC,Saltel F,Preville X,et al.Cathepsin-B-dependent apoptosis triggered by antithymocyte globulins:A novel mechanism of T-cell depletion[J].Blood.2003,102:3719-3726.
[56]Foghsgaard L,Wissing D,Mauch D,etal.Cathepsin B acts as a dominant execution protease in tumor cell apoptosis induced by tumor necrosis factor[J].J Cell Biol.2001,153(5):999-1010.
[57]Paris C,Bertoglio J,Bréard J.Lysosomal and mitochondrial pathways in miltefosine-induced apoptosis in U937 cells[J].Apoptosis.2007,12:1257-1267
[58]Nilsson E,Ghassemifar R,Brunk T.Lysosomal heterogeneity between and within cells with respect to resistance against oxidative stress[J].J Histochem,1997,29(11-12):857-865.
[59]Kagedal K,Zhao M,Svensson I,etal.Sphingosine-induced apoptosis is dependent on lysosomal proteases[J].Biochem J.2001,359(2):335-343.
[60]Bidere N,Lorenzo HK,Carmona S,etal.Cathepsin D triggers Bax activation,resulting in selective apoptosis-inducing factor(AIF) relocation in T lymphocytes entering the early commitment phase to apoptosis[J].J Biol Chem.2003,278(33):31401-31411.
[61]Heinrich M,Neumeyer J,Jakob M,et al.Cathepsin D links TNF-induced acid sphingomyelinase to Bid-mediated caspase-9 and -3 activation[J].Cell Death Differ,2004,11(5):550-563.
[62]Liu X,Kim CN,Yang J,et al.Induction of apoptotic program in cell-free extracts:Requirement for dATP and cytochrome c[J].Cell.1996,86(1):147-157.
[63]Susin SA,Lorenzo HK,Zamzami N,et al.Molecular characterization of mitochondrial apoptosis-inducing factor[J].Nature.1999,397(6718 ):441-446.
[64]Du C,Fang M,Li Y,et al.Smac,a mitochondrial protein that promotes cytochrome c-dependent caspase activation by eliminating IAP inhibition[J].Cell.2000,102(1):33-42.
[65]Verhagen AM,Ekert PG,Pakusch M,et al.Identification of DIABLO,a mammalian protein that promotes apoptosis by binding to and antagonizing IAP proteins[J].Cell.2000,102(1):43-53.
[66]Li LY,Luo X,Wang X.Endonuclease G(EndoG) is an apoptotic DNAse when released from mitochondria[J].Nature.2001,412(6842):95-99.
[67]Adams JM,Cory S.Life-or-death decisions by the Bcl-2 protein family[J].Trends Biochem Sci.2001,26(1):61-66.
[68]Cory S,Adams JM.The Bcl-2 family:regulators of the cellular life-or-death switch[J].Nat Rev Cancer.2002,2(9):647-656.
[69]Shih YL,Lin C J,Hsu SW,Cadmium toxicity toward caspase-independent apoptosis through the mitochondria-calcium pathway in mtDNA-depleted cells[J]Ann N Y Acad Sci.2005,1042:497-505
[70]Lemarié A,Lagadic-Gossmann D,Morzadec C.Cadmium induces caspase-independent apoptosis in liver Hep3B cells:role for calcium in signaling oxidative stress-related impairment of mitochondria and relocation of endonuclease G and apoptosis-inducing factor[J].Free Radic Biol Med.2004,36(12):1517-31
[71]Chen LB.Mitochondrial membrane potential in living cells.[J].Annu.Rev.cell Dev Biol.1988,4:155-181.
[72]Ye H,Cande C,Stephanou NC,et al.DNA binding is required for the apoptogenic action of apoptosis inducing factor[J].Nat.Struct.Biol.2002,9(9):680-685
[73]付玉荣,伊正君,颜玉蓉,等.HCPT诱导肝癌细胞凋亡时AIF定位变化与DNA损伤[J].肿瘤.2007,27(7):511-514
[74]Wang M,Zhang L,Han X,et al.Atiprimod inhibits the growth of mantle cell lymphoma in vitro and in vivo and induces apoptosis via activating the mitochondrial pathways[J].Blood 109:5455-5462
[75]Liou AK,Zhou Z,Pei W,et al.BimEL up-regulation potentiates AIF translocation and cell death in response to MPTP[J].FASEB.2005,19(10):1350-52
[76]Polster BM,Basanez G,Etxebarria A,et al.Calpain I Induces Cleavage and Release of Apoptosis-inducing Factor from Isolated Mitochondria[J].J Biol Chem.2005,280(8):6447-54.
[77]Ishihara I,Minami Y,Nishizaki T,et al.Activation of calpain precedes morphological alterations during hydrogen peroxide-induced apoptosis in neuronally differentiated mouse embryonal carcinoma P19 cell line[J].Neuroscience Letters.2000,279(2):97-100
[78]Kaiser N,Edelman IS.Calcium dependence of glucocorticoid-induced lymphocytolysis[J].PNAS.1977,74(2):638-642
[79] Weraeburg NW, Guicciardi ME, Bronk SF, et al. Tumor necrosis factor-alpha-associated lysosomal permeabilization is cathepsin B dependent[J]. Am J Physiol Gastrointest Liver Physiol. 2002,283: G947-G956.
[80] Heinrich M, Wickel M, Schneider-Brachert W, et al. Cathepsin D targeted by acid sphingomyelinase-derived ceramide[J]. EMBO. 1999,18: 5252-5263.
[81] Zdolsek J, Zhang H, Roberg K, etal. H_2O_2-mediated damage to lysosomal membranes of J-774 cells[J]. Free Radic Res Commun. 1993,18(2): 71-85
[82] Zhao M, Brunk UT, Eaton JW. Delayed oxidant-induced cell death involves activation of phospholipase A_2[J]. FEBS Lett 2001, 509: 399-404.
[83] Suzuki YJ, Forman HJ, Sevanian A. Oxidants as stimulators of signal transduction.[J]. Free Rad Biol Med. 1997,22: 269-285.
[84] Roberts LR, Kurosawa H, Bronk SF, et al. Cathepsin B contributes to bile salt-induced apoptosis of rat hepatocytes[J]. Gastroenterology. 1997,113: 1714-1726.
[85] Jones B, Roberts PJ, Faubion WA, et al. Cystatin A expression reduces bile salt-induced apoptosis in a rat hepatoma cell line[J]. Am J Physiol.1998, 275: 723-730
[86]. Faubion WA, Guicciardi ME, Miyoshi H, et al. Toxic bile salts induce rodent hepatocyte apoptosis via direct activation of Fas[J]. J Clin Invest. 1999,103: 137-45.
[87] Faubion WA, Gores GJ. Death receptors in liver biology and pathobiology[J]. Hepatology. 1999, 29: 1-4.
[88] Wille A, Gerber A, Heimburg A, et al. Cathepsin L is involved in cathepsin D processing and regulation of apoptosis in A549 human lung epithelial cells[J]. Biol Chem. 2004, 385: 665-670.
[89] Baud O, Li J, Zhang Y, et al. Nitric oxide-induced cell death in developing oligodendrocytes is associated with mitochondrial dysfunction and apoptosis-inducing factor translocation[J]. Eur J Neurosci. 2004, 7(20): 1713-1726
[90] Averna M, Stifanese R, De Tullio R, et al. Regulation of calpain activity in rat brain with altered Ca~(2+) homeostasis[J]. J Biol Chem. 2007,282(4): 2656-65
[1]左伋.医学分子细胞生物学[M].上海:复巨大学出版社,2005:152-156
[2]Nagaraj NS,Vigneswaran N,Zacharias W.Hypoxia inhibits TRAIL-induced tumor cell apoptosis:Involvement of lysosomal cathepsins[J]Apoptosis.2007,12:125-139
[3]Joseph A.C,Patricia A.M,Aby J.Aryl Hydrocarbon Receptor Modulation of Tumor Necrosis Factor-induced Apoptosis and Lysosomal Disruption in a Hepatoma Model That Is Caspase-8-independent[J]J Biological Chem.2006,281(16):10954-10967
[4]De Duve C.Lysosomes revisited[J].Eur.J.Biochem.1983,137:391-397.
[5]Li W,Yuan XM,Nordgren G,et al.Induction of cell death by the lysosomotropic detergent MSDH[J].FEBS Lett.2000,470:35 -39
[6]Wilson PD,Firestone RA,Lenard J.The role of lysosomal enzymes in killing of mammalian cells by the lysosomotrpic detergent Ndodecylimidazole[J]J Cell Biol.1987,104:1223-1229
[7]Neuzil J,Svensson I,Weber T,et al.alpha-Tocopheryl succinate-induced apoptosis in Jurkat T cells involves caspase-3 activation,and both lysosomal and mitochondrial destabilisation[J].FEBS Lett.1999,445:295-300
[8]Neuzil J,Weber T,Schroder A,etal.Induction of cancer cell apoptosis by alpha-tocopherol succinate:molecular pathways and structural requirements[J].FASEB.2001,15:403-415
[9]Yuan X.M,Li W,Dalen H.,Lysosomal destabilization in p53-induced apoptosis.[J].PNAS.2002,99:6286-6291
[10]Ferri KF,Kroemer G.Organelle-specific initiation of cell death pathways[J].Nat Cell Biol.2001,3:E255-E263.
[11]Brunk UT,Neuzil J,Eaton JW.Lysosomal involvement in apoptosis[J].Redox Rep.2001,6:91-97.
[12]Kagedal K,ZhaoM,Svensson I,et al.phingosine-induced apoptosis is dependent on lysosomal proteases[J].Biochem J.2001,359:335-343.
[13]Werneburg NW,Guicciardi ME,Bronk SF,et al.Tumor necrosis factor-alpha-associated lysosomal permeabilization is cathepsin B dependent[J].Am J Physiol Gastrointest Liver Physiol.2002,283:G947-G956.
[14]Heinrich M,Wickel M,SchneiderBW,et al.Cathepsin D targeted by acid sphingomyelinase-derived ceramide[J].EMBO.1999,18:5252-5263.
[15]Guicciardi MF,Bronk SF,Wemeburg NW,et al.cFLIPL prevents TRAIL-induced apoptosis of hepatocellular carcinoma cells by inhibiting the lysosomal pathway of apoptosis[J].Am J Physiol Gastrointest Liver Physiol.2007,292:G 1337-G1346.
[16]Guicciardi ME,Leist M,Gores GJ.Lysosomes in cell death.Oncogene[J].2004,23: 2881-2890.
[17] Caroline P, Jacques B, Jacqueline B. Lysosomal and mitochondrial pathways in miltefosine-induced apoptosis in U937 cells[J]. Apoptosis. 2007,12:1257-1267
[18] Paquet C, Sane AT, Beauchemin M, et al. Caspase and mitochondrial dysfunction-dependent mechanisms of lysosomal eakage and cathepsin B activation in DNA damage-induced apoptosis[J]. Leukemia.2005,19: 784-791
[19] Kagedal K, Johansson AC, Johansson U, et al. Lysosomal membrane permeabilization during apoptosis—involvement of Bax?[J]. Int J Exp Patho. 2005,186:309-321
[20] Zdolsek J, Zhang H, Roberg K, Brunk U. H_2O_2-mediated damage to lysosomal membranes of J-774 cells[J] Free Radic Res Commun. 1993, 18: 71-85.
[21] Antunes F, Cadenas E, Brunk UT. Apoptosis induced by exposure to a low steady-state concentration of H_2O_2 is a consequence of lysosomal rupture[J].Biochem J. 2001, 356: 549-555.
[22] Persson HL, Yu Z, Tirosh O, et al. Prevention of oxidant-induced cell death by lysosomotropic iron chelators[J]. Free Radical Biol Med. 2003,34: 1295-1305.
[23] Zhao M, Brunk UT, Eaton JW. Delayed oxidant-induced cell death involves activation of phospholipase A2[J]. FEBS Lett. 2001, 509: 399-404.
[24] Zhao M, Antunes F, Eaton JW, Brunk UT. Lysosomal enzymes promote mitochondrial oxidant production, cytochrome c release and apoptosis[J]. Eur J Biochem. 2003,270: 3778-3786.
[25] Suzuki YJ, Forman HJ, Sevanian A. Oxidants as stimulators of signal transduction[J]. Free Rad Biol Med. 1997, 22: 269-285.
[26] Yap YW, Whiteman M, Bay BH, et al. Hypochlorous acid induces apoptosis of cultured cortical neurons through activation of calpains and rupture of lysosomes[J]. J Neurochem.. 2006, 98: 1597-1609
[27] Nylandsted J, M. Gyrd-Hansen, A. Danielewicz, N,etal. Heat shock protein 70 promotes cell survival by inhibiting lysosomal membrane permeabilization[J]. J. Exp. Med. 2004, 200: 425-435.
[28] Nylandsted J, Wick W, Hirt UA, et al. Eradication of glioblastoma, and breast and colon carcinoma xenografts by Hsp70 depletion[J]. Cancer Res. 2002, 62: 7139-7142.
[29] Michallet MC, Saltel F, Preville X, et al. Cathepsin-B-dependent apoptosis triggered by antithymocyte globulins: A novel mechanism of T-cell depletion[J]. Blood. 2003,102: 3719-3726.
[30] Michallet MC, Saltel F, Flacher M, et al. Cathepsin-dependent apoptosis triggered by supraoptimal activation of T lymphocytes: A possible mechanism of high dose tolerance[J]. J Immunol 2004; 172: 5405-5414.
[31]Bidere N,LorenzoHK,Carmona S,et al.CathepsinD triggers Bax activation,resulting in selective apoptosis-inducing factor(AIF) relocation in T lymphocytes entering the early commitment phase to apoptosis[J].J Biological Chem.2003,278:31401-31411.
[32]Vancompetnolle K,VanHetreweghe F,Pynaert G,et al.Atmcty los ide-induced release of cathepsin B,a protease with caspase-processing activity[J].FEES Lett.1998,438:150-158
[33]Zhao M,Eaton JW,Brunk UT.Protection against oxtdart-mediated lysosomal rupture:a new anti-apoptotic activity of Bcl-2[J].FEBS Lett.2000,485:104-108
[34]Ishisaka R,Utsumi T,Yabuki M,et al.Activation of caspase-3 Like protease by digitonin-treated lysosomes[J].FEBS Lett.1998,435:233-236
[35]Chwieralski CE,Welte T,Bühling F.Cathepsin-regulated apoptosis[J].Apoptosis.2006,11:143-149
[36]Turk B,Turk D,Turk V.Lysosomal cysteine proteases:more than scavengers[J].Biochim Biophys Acta.2000,1477:98-111
[37]Leist M,Jaattela M.Triggering of apoptosis by cathepsins[J].Cell Death Differ.2001,8:324-326
[38]Turk V,Turk B,Turk D.Lysosomal cysteine proteases:facts and opportunities[J].EMBO J.2001,20:4629-06
[39]Taha TA,Kitatani K,Bielawski J,et al.TNF induces the loss of sphingosine kinase-1 by a cathepsin B dependent mechanism[J].J.Biol.Chem.2005,280:17196-17202.
[40]Guicciardi ME,Deussing J,Miyoshi H,et al.Cathepsin B contributes to TNF-alpha-mediated hepatocyte apoptosis by promoting mitochondrial release of cytochrome c[J].J Clin Invest.2000;106:1127-1137.
[41]Tang PS,Tsang ME,Lodyga M,et al.Lipopolysaccharide Accelerates Caspase-Independent but Cathepsin B-Dependent Death of Human Lung Epithelial Cells[J].J.Cell Physiol.2006,209:457-467
[42]Nagaraj NS,Vigneswaran N,Zacharias W.Cathepsin B mediates TRAIL-induced apoptosis in oral cancer cells[J].J Cancer Res Clin Oncol.2006,132:171-183
[43]Li JH,Pober JS.The Cathepsin B Death Pathway Contributes to TNF Plus IFN-Mediated Human Endothelial Injury[J].J.Immunol.2005,175:1858-1866.
[44]Liu N,Raja SM,Zazzeroni F,et al.NF-kappaB protects from the lysosomal pathway of cell death[J].EMBO J.2003,22:5313-5322.
[45]Canbay A,Guicciardi ME,Higuchi H,et al.Cathepsin B inactivation attenuates hepatic injury and fibrosis during cholestasis[J].J Clin Invest.2003,112:152-159.
[46]Roberts LR,Kurosawa H,Bronk SF,et al.Cathepsin B contributes to bile salt-induced apoptosis of rat hepatocytes[J]. Gastroenterology. 1997, 113: 1714-1726.
[47] Jones B, Roberts PJ, FaubionWA, et al. Cystatin A expression reduces bile salt-induced apoptosis in a rat hepatoma cell line[J]. Am J Physiol. 1998, 275: G723-G730.
[48] Faubion WA, Guicciardi ME, Miyoshi H, et al. Toxic bile salts induce rodent hepatocyte apoptosis via direct activation of Fas[J]. J Clin Invest. 1999,103: 137-145.
[49] Faubion WA, Gores GJ. Death receptors in liver biology and pathobiology[J]. Hepatology. 1999;29:1-4.
[50] Salvesen G. A Iysosomal protease enters the death scene[J] J Clin Invest. 2001, 107:21-22.
[51] Stoka V, Turk B, Schendel SL, et al. Lysosomal protease pathways to apoptosis: cleavage of bid, not Pro-caspases, is the most likely route[J]. J Biological Chem. 2000, 276: 3149-3157.
[52] Houseweart MK, Vilaythong A, Yin XM, et al. Apoptosis caused by cathepsins does not require Bid signaling in an in vivo model of progressive myoclonus epilepsy (EPM1) [J]. Cell Death Differ. 2003,10: 1329-1335.
[53] Vancompernolle K, Van H F, Pynaert G, et al. Atractyloside-induced release of cathepsin B, a protease with caspase-processing activity[J]. FEBS Lett. 1998, 438: 150-158.
[54] Bro"ker LE., Huisman C, Span SW, et al. Cathepsin B Mediates Caspase-Independent Cell Death Induced by Microtubule Stabilizing Agents in Non-Small Cell Lung Cancer Cells[J]. Cancer Res. 2004, 64: 27-30.
[55] Felbor U, Kessler B, Mothes W, et al. Neuronal loss and brain atrophy in mice lacking cathepsins B and L[J]. Proc Natl Acad Sci. 2002, 99: 7883-7888
[56] Heinrich M, Neumeyer J, Jakob M, et al. Cathepsin D links TNF-induced acid sphingomyelinase to Bid-mediated caspase-9 and -3 activation[J]. Cell Death Differ, 2004, 11(5):550-563.
[57] Wille A, Gerber A, Heimburg A, et al. Cathepsin L is involved in cathepsin D processing and regulation of apoptosis in A549 human lung epithelial cells[J]. Biol Chem. 2004, 385: 665-670.
[58] Tardy C, Codogno P, Autefage H, et al. Lysosomes and lysosomal proteins in cancer celldeath[J]. Biochem Biophys Acta. 2006,1765: 101-125
