Bcl-2 associated athanogene 5 (Bag5) is overexpressed in prostate cancer and inhibits ER-stress induced apoptosis

BMC Cancer

Anja Bruchmann²  Corinna Roller²  Tamara Vanessa Walther²  Georg Schäfer¹  Sara Lehmusvaara³  Tapio Visakorpi³  Helmut Klocker¹  Andrew C B Cato²  Danilo Maddalo² 

[1] Division of Experimental Urology, Department of Urology, Innsbruck Medical University, Innsbruck, Austria

[2] Karlsruhe Institute of Technology, Institute of Toxicology and Genetics, Hermann-von-Helmholtz Platz 1, Eggenstein-Leopoldshafen 76344, Germany

[3] Institute of Biomedical Technology and BioMediTech, University of Tampere and Tampere University Hospital, Tampere FI-33014, Finland

关键词: Refolding;    Molecular chaperones;    Apoptosis;    Endoplasmic reticulum;    Cell stress;    Unfolded protein response;   

Others  :  1079894
DOI  :  10.1186/1471-2407-13-96

 received in 2012-08-03, accepted in 2013-02-18,  发布年份 2013

【 摘 要 】

Background

The Bag (Bcl-2 associated athanogene) family of proteins consists of 6 members sharing a common, single-copied Bag domain through which they interact with the molecular chaperone Hsp70. Bag5 represents an exception in the Bag family since it consists of 5 Bag domains covering the whole protein. Bag proteins like Bag1 and Bag3 have been implicated in tumor growth and survival but it is not known whether Bag5 also exhibits this function.

Methods

Bag5 mRNA and protein expression levels were investigated in prostate cancer patient samples using real-time PCR and immunoblot analyses. In addition immunohistological studies were carried out to determine the expression of Bag5 in tissue arrays. Analysis of Bag5 gene expression was carried out using one-way ANOVA and Bonferroni’s Multiple Comparison test. The mean values of the Bag5 stained cells in the tissue array was analyzed by Mann-Whitney test. Functional studies of the role of Bag5 in prostate cancer cell lines was performed using overexpression and RNA interference analyses.

Results

Our results show that Bag5 is overexpressed in malignant prostate tissue compared to benign samples. In addition we could show that Bag5 levels are increased following endoplasmic reticulum (ER)-stress induction, and Bag5 relocates from the cytoplasm to the ER during this process. We also demonstrate that Bag5 interacts with the ER-resident chaperone GRP78/BiP and enhances its ATPase activity. Bag5 overexpression in 22Rv.1 prostate cancer cells inhibited ER-stress induced apoptosis in the unfolded protein response by suppressing PERK-eIF2-ATF4 activity while enhancing the IRE1-Xbp1 axis of this pathway. Cells expressing high levels of Bag5 showed reduced sensitivity to apoptosis induced by different agents while Bag5 downregulation resulted in increased stress-induced cell death.

Conclusions

We have therefore shown that Bag5 is overexpressed in prostate cancer and plays a role in ER-stress induced apoptosis. Furthermore we have identified GRP78/BiP as a novel interaction partner of Bag5.

【 授权许可】

   
2013 Bruchmann et al; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Doong H, Vrailas A, Kohn EC: What’s in the ‘BAG’?–A functional domain analysis of the BAG-family proteins. Cancer Lett 2002, 188(1–2):25-32.
• [2]Sondermann H, Scheufler C, Schneider C, Hohfeld J, Hartl FU, Moarefi I: Structure of a Bag/Hsc70 complex: convergent functional evolution of Hsp70 nucleotide exchange factors. Science 2001, 291(5508):1553-1557.
• [3]Briknarova K, Takayama S, Brive L, Havert ML, Knee DA, Velasco J, Homma S, Cabezas E, Stuart J, Hoyt DW: Structural analysis of BAG1 cochaperone and its interactions with Hsc70 heat shock protein. Nat Struct Biol 2001, 8(4):349-352.
• [4]Brimmell M, Burns JS, Munson P, McDonald L, O’Hare MJ, Lakhani SR, Packham G: High level expression of differentially localized BAG-1 isoforms in some oestrogen receptor-positive human breast cancers. Br J cancer 1999, 81(6):1042-1051.
• [5]Briknarova K, Takayama S, Homma S, Baker K, Cabezas E, Hoyt DW, Li Z, Satterthwait AC, Ely KR: BAG4/SODD protein contains a short BAG domain. J Biol Chem 2002, 277(34):31172-31178.
• [6]Takayama S, Sato T, Krajewski S, Kochel K, Irie S, Millan JA, Reed JC: Cloning and functional analysis of BAG-1: a novel Bcl-2-binding protein with anti-cell death activity. Cell 1995, 80(2):279-284.
• [7]Wang HG, Reed JC: Bc1-2, Raf-1 and mitochondrial regulation of apoptosis. Biofactors 1998, 8(1–2):13-16.
• [8]Lee JH, Takahashi T, Yasuhara N, Inazawa J, Kamada S, Tsujimoto Y: Bis, a Bcl-2-binding protein that synergizes with Bcl-2 in preventing cell death. Oncogene 1999, 18(46):6183-6190.
• [9]Jiang Y, Woronicz JD, Liu W, Goeddel DV: Prevention of constitutive TNF receptor 1 signaling by silencer of death domains. Science 1999, 283(5401):543-546.
• [10]Miki K, Eddy EM: Tumor necrosis factor receptor 1 is an ATPase regulated by silencer of death domain. Mol Cell Biol 2002, 22(8):2536-2543.
• [11]Thress K, Henzel W, Shillinglaw W, Kornbluth S: Scythe: a novel reaper-binding apoptotic regulator. EMBO J 1998, 17(21):6135-6143.
• [12]Thress K, Song J, Morimoto RI, Kornbluth S: Reversible inhibition of Hsp70 chaperone function by Scythe and Reaper. EMBO J 2001, 20(5):1033-1041.
• [13]Andre B, Springael JY: WWP, a new amino acid motif present in single or multiple copies in various proteins including dystrophin and the SH3-binding Yes-associated protein YAP65. Biochem Biophys Res Commun 1994, 205(2):1201-1205.
• [14]Doong H, Price J, Kim YS, Gasbarre C, Probst J, Liotta LA, Blanchette J, Rizzo K, Kohn EC: CAIR-1/BAG-3 forms an EGF-regulated ternary complex with phospholipase C-γ and Hsp70/Hsc70. Oncogene 2000, 19(38):4385-4395.
• [15]Manchen ST, Hubberstey AV: Human Scythe contains a functional nuclear localization sequence and remains in the nucleus during staurosporine-induced apoptosis. Biochem Biophys Res Commun 2001, 287:1075-1082.
• [16]Townsend PA, Curtess RI, Sharp A, Brimmel M, Packham GK: BAG-1: a multifunctional regulator of cell growth and survival. Biochimica and Biophysica Acta 2003, 1603:83-98.
• [17]Kermer P, Digicaylioglu MH, Kaul M, Zapata JM, Krajewska M, Stenner-Liewen F, Takayama S, Krajewski S, Lipton SA, Reed JC: BAG1 over-expression in brain protects against stroke. Brain Pathol 2003, 13(4):495-506.
• [18]Annunziata CM, Kleinberg L, Davidson B, Berner A, Gius D, Tchabo N, Steinberg SM, Kohn EC: BAG-4/SODD and associated antiapoptotic proteins are linked to aggressiveness of epithelial ovarian cancer. Clin Cancer Res 2007, 13(22 Pt 1):6585-6592.
• [19]Bonelli P, Petrella A, Rosati A, Romano MF, Lerose R, Pagliuca MG, Amelio T, Festa M, Martire G, Venuta S: BAG3 protein regulates stress-induced apoptosis in normal and neoplastic leukocytes. Leukemia 2004, 18(2):358-360.
• [20]Liao Q, Ozawa F, Friess H, Zimmermann A, Takayama S, Reed JC, Kleeff J, Buchler MW: The anti-apoptotic protein BAG-3 is overexpressed in pancreatic cancer and induced by heat stress in pancreatic cancer cell lines. FEBS Lett 2001, 503(2–3):151-157.
• [21]Chung KK, Dawson TM: Parkin and Hsp70 sacked by BAG5. Neuron 2004, 44(6):899-901.
• [22]Wang X, Guo J, Jiang H, Shen L, Tang B: Direct interaction between BAG5 protein and Parkin protein. Zhong Nan Da Xue Xue Bao Yi Xue Ban 2010, 35(11):1156-1161.
• [23]Bartsch G, Horninger W, Klocker H, Pelzer A, Bektic J, Oberaigner W, Schennach H, Schafer G, Frauscher F, Boniol M: Tyrol Prostate Cancer Demonstration Project: early detection, treatment, outcome, incidence and mortality. BJU Int 2008, 101(7):809-816.
• [24]Oberaigner W, Siebert U, Horninger W, Klocker H, Bektic J, Schafer G, Frauscher F, Schennach H, Bartsch G: Prostate-specific antigen testing in Tyrol, Austria: prostate cancer mortality reduction was supported by an update with mortality data up to 2008. Int J Public Health 2012, 57(1):57-62.
• [25]Kong L, Schafer G, Bu H, Zhang Y, Klocker H: Lamin A/C protein is overexpressed in tissue-invading prostate cancer and promotes prostate cancer cell growth, migration and invasion through the PI3K/AKT/PTEN pathway. Carcinogenesis 2012, 33(4):751-759.
• [26]Webber MM, Quader ST, Kleinman HK, Bello-DeOcampo D, Storto PD, Bice G, DeMendonca-Calaca W, Williams DE: Human cell lines as an in vitro/in vivo model for prostate carcinogenesis and progression. Prostate 2001, 47(1):1-13.
• [27]Townsend PA, Cutress RI, Sharp A, Brimmell M, Packham G: BAG-1 prevents stress-induced long-term growth inhibition in breast cancer cells via a chaperone-dependent pathway. Cancer Res 2003, 63(14):4150-4157.
• [28]Lai E, Teodoro T, Volchuk A: Endoplasmic reticulum stress: signaling the unfolded protein response. Physiology (Bethesda) 2007, 22:193-201.
• [29]Szegezdi E, Logue SE, Gorman AM, Samali A: Mediators of endoplasmic reticulum stress-induced apoptosis. EMBO Rep 2006, 7(9):880-885.
• [30]Li J, Ni M, Lee B, Barron E, Hinton DR, Lee AS: The unfolded protein response regulator GRP78/BiP is required for endoplasmic reticulum integrity and stress-induced autophagy in mammalian cells. Cell Death Differ 2008, 15(9):1460-1471.
• [31]Lin JH, Li H, Yasumura D, Cohen HR, Zhang C, Panning B, Shokat KM, Lavail MM, Walter P: IRE1 signaling affects cell fate during the unfolded protein response. Science 2007, 318(5852):944-949.
• [32]Lin JH, Li H, Zhang Y, Ron D, Walter P: Divergent effects of PERK and IRE1 signaling on cell viability. PLoS One 2009, 4(1):e4170.
• [33]McCullough KD, Martindale JL, Klotz LO, Aw TY, Holbrook NJ: Gadd153 sensitizes cells to endoplasmic reticulum stress by down-regulating Bcl2 and perturbing the cellular redox state. Mol Cell Biol 2001, 21(4):1249-1259.
• [34]Maddalo D, Neeb A, Jehle K, Schmitz K, Muhle-Goll C, Shatkina L, Walther TV, Bruchmann A, Gopal SM, Wenzel W: A peptidic unconjugated GRP78/BiP ligand modulates the unfolded protein response and induces prostate cancer cell death. PLoS One 2012, 7(10):e45690.
• [35]Ammirante M, De Laurenzi V, Graziano V, Turco MC, Rosati A: BAG3 is required for IKKalpha nuclear translocation and emergence of castration resistant prostate cancer. Cell Death Dis 2011, 2:e139.
• [36]Krajewska M, Turner BC, Shabaik A, Krajewski S, Reed JC: Expression of BAG-1 protein correlates with aggressive behavior of prostate cancers. Prostate 2006, 66(8):801-810.
• [37]Maki HE, Saramaki OR, Shatkina L, Martikainen PM, Tammela TL, van Weerden WM, Vessella RL, Cato AC, Visakorpi T: Overexpression and gene amplification of BAG-1L in hormone-refractory prostate cancer. J Pathol 2007, 212(4):395-401.
• [38]Townsend PA, Dublin E, Hart IR, Kao RH, Hanby AM, Cutress RI, Poulsom R, Ryder K, Barnes DM, Packham G: BAG-1 expression in human breast cancer: interrelationship between BAG-1 RNA, protein, HSC70 expression and clinico-pathological data. J Pathol 2002, 197(1):51-59.
• [39]Yang X, Hao Y, Ding Z, Pater A, Tang SC: Differential expression of antiapoptotic gene BAG-1 in human breast normal and cancer cell lines and tissues. Clinical cancer research: an official journal of the American Association for Cancer Research 1999, 5(7):1816-1822.
• [40]Clemo NK, Collard TJ, Southern SL, Edwards KD, Moorghen M, Packham G, Hague A, Paraskeva C, Williams AC: BAG-1 is up-regulated in colorectal tumour progression and promotes colorectal tumour cell survival through increased NF-kappaB activity. Carcinogenesis 2008, 29(4):849-857.
• [41]Ozawa F, Friess H, Zimmermann A, Kleeff J, Buchler MW: Enhanced expression of Silencer of death domains (SODD/BAG-4) in pancreatic cancer. Biochem Biophys Res Commun 2000, 271(2):409-413.
• [42]Liao Q, Ozawa F, Friess H, Zimmermann A, Takayama S, Reed JC, Kleeff J, Buchler MW: The anti-apoptotic protein BAG-3 is overexpressed in pancreatic cancer and induced by heat stress in pancreatic cancer cell lines. FEBS letters 2001, 503(2–3):151-157.
• [43]Chou JW, Bushel PR: Discernment of possible mechanisms of hepatotoxicity via biological processes over-represented by co-expressed genes. BMC Genomics 2009, 10:272. BioMed Central Full Text
• [44]Hernandez-Vargas H, von Kobbe C, Sanchez-Estevez C, Julian-Tendero M, Palacios J, Moreno-Bueno G: Inhibition of paclitaxel-induced proteasome activation influences paclitaxel cytotoxicity in breast cancer cells in a sequence-dependent manner. Cell Cycle 2007, 6(21):2662-2668.
• [45]L’Esperance S, Bachvarova M, Tetu B, Mes-Masson AM, Bachvarov D: Global gene expression analysis of early response to chemotherapy treatment in ovarian cancer spheroids. BMC Genomics 2008, 9:99. BioMed Central Full Text
• [46]Roller C, Maddalo D: The molecular chaperone GRP78/BiP in the development of chemoresistance: mechanism and possible treatment. Frontiers Pharmacology 2013, 4:10.