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BAG-1 suppresses expression of the key regulatory cytokine transforming growth factor β (TGF-β1) in colorectal tumour cells

Oncogene volume 32, pages 4490–4499 (2013)Cite this article

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Abstract

As colorectal cancer remains the second highest cause of cancer-related deaths in much of the industrialised world, identifying novel strategies to prevent colorectal tumour development remains an important challenge. BAG-1 is a multi-functional protein, the expression of which is up-regulated at relatively early stages in colorectal tumorigenesis. Importantly, BAG-1 is thought to enhance colorectal tumour progression through promoting tumour cell survival. Here, we report for the first time a novel role for BAG-1, establishing it as a suppressor of transforming growth factor β (TGF-β1) expression in colorectal tumour cells. Microarray analysis first highlighted the possibility that BAG-1 may regulate TGF-β1 expression, a key cytokine in normal colonic tissue homoeostasis. Q-RT–PCR and ELISA demonstrated TGFB1 mRNA and protein expression to be significantly increased when BAG1 levels were reduced by small interfering RNA; additionally, induction of BAG-1L caused suppression of TGFB1 mRNA in colorectal tumour cells. Using reporter and chromatin immunoprecipitation assays, a direct association of BAG-1 with the TGFB1 gene regulatory region was identified. Immunohistochemistry and Weiser fraction data indicated that the levels of BAG-1 and TGF-β1 are inversely correlated in the normal colonic epithelium in vivo, consistent with a role for BAG-1-mediated repression of TGF-β1 production. In vitro studies showed that the change in TGF-β1 production following manipulation of BAG-1 is functionally relevant; through induction of anchorage-independent growth in TGF-β1-dependent normal rat kidney fibroblasts and regulation of SMAD2 phosphorylation in TGF-β1-sensitive adenoma cells. Taken together, this study identifies the anti-apoptotic protein BAG-1 as a suppressor of the inhibitory growth factor TGF-β1, suggesting that high expression of BAG-1 can impact on a number of the hallmarks of cancer, of potential importance in promoting the early stages of colorectal tumorigenesis. Establishing BAG-1 as a repressor of TGF-β1 has important biological implications, and highlights a new role for BAG-1 in colorectal tumorigenesis.

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References

  1. Massagué J . TGFbeta in cancer. Cell 2008; 134: 215–230.

    Article  Google Scholar 

  2. Ikushima H, Miyazono K . TGF-beta signalling: a complex web in cancer progression. Nat Rev Cancer 2010; 10: 415–424.

    Article  CAS  Google Scholar 

  3. Moustakas A, Heldin CH . The regulation of TGFbeta signal transduction. Development 2009; 136: 3699–3714.

    Article  CAS  Google Scholar 

  4. Avery A, Paraskeva C, Hall P, Flanders KC, Sport M, Moorghen M . TGF-beta expression in the human colon: differential immunostaining along the crypt epithelium. Br J Cancer 1993; 68: 137–139.

    Article  CAS  Google Scholar 

  5. Barnard JA, Warwick GJ, Gold LI . Localization of transforming growth factor beta isoforms in the normal murine small intestine and colon. Gastroenterology 1993; 105: 67–73.

    Article  CAS  Google Scholar 

  6. Heldin CH, Landström M, Moustakas A . Mechanism of TGF-beta signalling to growth arrest, apoptosis and epithelial-mesenchymal transition. Curr Opin Cell Biol 2009; 21: 166–176.

    Article  CAS  Google Scholar 

  7. Lampropoulos P, Zizi-Sermpetzoglou A, Rizos S, Kostakis A, Nikiteas N, Papavassiliou AG . TGF-beta signalling in colon carcinogenesis. Cancer Lett 2012; 314: 1–7.

    Article  CAS  Google Scholar 

  8. Babyatsky MW, Rossiter G, Podolsky DK . Expression of transforming growth factors alpha and beta in colonic mucosa in inflammatory bowel disease. Gastroenterology 1996; 110: 798–802.

    Google Scholar 

  9. Townsend PA, Stephanou A, Packham G, Latchman DS . BAG-1: a multi-functional pro-survival protein. Int J Biochem Cell Biol 2005; 37: 251–259.

    Article  CAS  Google Scholar 

  10. Packham G, Brimmell M, Cleveland JL . Mammalian cells express two differently localized Bag-1 isoforms generated by alternative translation initiation. Biochem J 1997; 328: 807–813.

    Article  CAS  Google Scholar 

  11. Sharp A, Crabb SJ, Townsend PA, Cutress RI, Brimmell M, Wang XH et al. BAG-1 in carcinogenesis. Exp Rev Mol Med 2004; 6: 1–15.

    Article  Google Scholar 

  12. Clemo NK, Collard TJ, Southern SL, Edwards KD, Moorghen M, Packham G et al. BAG-1 is up-regulated in colorectal tumour progression and promotes colorectal tumour cell survival through increased NF-kappaB activity. Carcinogenesis 2008; 29: 849–857.

    Article  CAS  Google Scholar 

  13. Arhel NJ, Packham G, Townsend PA, Collard TF, H-Zadeh AM, Sharp A et al. The retinoblastoma protein interacts with Bag-1 in human colonic adenoma and carcinoma derived cell lines. Int J Cancer 2003; 106: 364–371.

    Article  CAS  Google Scholar 

  14. Barnes JD, Arhel NJ, Lee SS, Sharp A, Al-Okail M, Packham G et al. Nuclear BAG-1 expression inhibits apoptosis in colorectal adenoma-derived epithelial cells. Apoptosis 2005; 10: 301–311.

    Article  CAS  Google Scholar 

  15. Kikuchi R, Noguchi T, Takeno S, Funada Y, Moriyama H, Uchida Y . Nuclear BAG-1 expression reflects malignant potential in colorectal carcinomas. Br J Cancer 2002; 87: 1136–1139.

    Article  CAS  Google Scholar 

  16. Gehring U . Multiple, but concerted cellular activities of the human protein Hap46/BAG-1M and isoforms. Int J Mol Sci 2009; 10: 906–928.

    Article  CAS  Google Scholar 

  17. Niyaz Y, Zeiner M, Gehring U . Transcriptional activation by human Hsp70-associating protein Hap50. J Cell Sci 2001; 114: 1839–1845.

    CAS  Google Scholar 

  18. Zeiner M, Niyaz Y, Gehring U . The hsp70-associating protein Hap46 binds to DNA and stimulates transcription. Proc Natl Acad Sci USA 1999; 96: 10194–10199.

    Article  CAS  Google Scholar 

  19. Cutress RI, Townsend PA, Sharp A, Maison A, Wood L, Lee R et al. The nuclear BAG-1 isoform, BAG-1L, enhances oestrogen-dependent transcription. Oncogene 2003; 22: 4973–4982.

    Article  CAS  Google Scholar 

  20. Shatkina L, Mink S, Rogatsch H, Klocker H, Langer G, Nestl A et al. The cochaperone Bag-1L enhances androgen receptor action via interaction with the NH2-terminal region of the receptor. Mol Cell Biol 2003; 23: 7189–7197.

    Article  CAS  Google Scholar 

  21. Wang XH, O’Connor D, Brimmell M, Packham G . The BAG-1 co-chaperone is a negative regulator of p73-dependent transcription. Br J Cancer 2009; 100: 1347–1357.

    Article  CAS  Google Scholar 

  22. Southern SL, Collard TJ, Urban BC, Skeen VR, Smartt HJ, Hague A et al. BAG-1 interacts with the p50-p50 homodimeric NF-kappaB complex: implications for colorectal carcinogenesis. Oncogene 2012; 31: 2761–2772.

    Article  CAS  Google Scholar 

  23. Takayama S, Krajewski S, Krajewska M, Kitada S, Zapata JM, Kochel K et al. Expression and location of Hsp70/Hsc-binding anti-apoptotic protein BAG-1 and its variants in normal tissues and tumour cell lines. Cancer Res 1998; 58: 3116–3131.

    CAS  Google Scholar 

  24. Kim SJ, Glick A, Sporn MB, Roberts AB . Characterization of the promoter region of the human transforming growth factor-beta 1 gene. J Biol Chem 1989; 264: 402–408.

    CAS  PubMed  Google Scholar 

  25. Barnard JA, Warwick GJ . Butyrate rapidly induces growth inhibition and differentiation in HT-29 cells. Cell Growth Diff 1993; 4: 495–501.

    CAS  PubMed  Google Scholar 

  26. Wang G, Higgins PJ, Gannon M, Staiano-Coico L . Transforming growth factor-beta 1 acts cooperatively with sodium n-butyrate to induce differentiation of normal human keratinocytes. Exp Cell Res 1992; 198: 27–30.

    Article  CAS  Google Scholar 

  27. Hague A, Singh B, Paraskeva C . Butyrate acts as a survival factor for colonic epithelial cells: further fuel for the in vivo versus in vitro debate. Gastroenterology 1997; 112: 1036–1040.

    Article  CAS  Google Scholar 

  28. Mariadason JM, Nicholas C, L’Italien KE, Zhuang M, Smartt HJ, Heerdt BG et al. Gene expression profiling of intestinal epithelial cell maturation along the crypt-villus axis. Gastroenterology 2005; 128: 1081–1088.

    Article  CAS  Google Scholar 

  29. Smartt HJ, Greenhough A, Ordóñez-Morán P, Talero E, Cherry CA, Wallam CA et al. Beta-catenin represses expression of the tumour suppressor 15-prostaglandin dehydrogenase in the normal intestinal epithelium and colorectal tumour cells. Gut 2012; 61: 1306–1314.

    Article  CAS  Google Scholar 

  30. Manning AM, Williams AC, Game SM, Paraskeva C . Differential sensitivity of human colonic adenoma and carcinoma cells to transforming growth factor beta (TGF-beta): conversion of an adenoma cell line to a tumorigenic phenotype is accompanied by a reduced response to the inhibitory effects of TGF-beta. Oncogene 1991; 6: 1471–1476.

    CAS  PubMed  Google Scholar 

  31. Skeen VR, Paterson I, Paraskeva C, Williams AC . TGF-β1 signalling, connecting aberrant inflammation and colorectal tumori-genesis. Curr Pharm Des 2012; 18: 3874–3888.

    Article  CAS  Google Scholar 

  32. Datto MB, Li Y, Panus JF, Howe DJ, Xiong Y, Wang XF . Transforming growth factor-beta induces the cyclin-dependent kinase inhibitor p21 through a p53-independent mechanism. Proc Natl Acad Sci USA 1995; 92: 5545–5549.

    Article  CAS  Google Scholar 

  33. Polyak K, Kato JY, Solomon MJ, Sherr CJ, Massagué J, Roberts JM et al. p27Kip1, a cyclin-Cdk inhibitor, links transforming growth factor-beta and contact inhibition to cell cycle arrest. Genes Dev 1994; 8: 9–22.

    Article  CAS  Google Scholar 

  34. Pardali K, Kurisaki A, Morén A, ten Dijke P, Kardassis D, Moustakas A . Role of Smad proteins and transcription factor Sp1 in p21(Waf1/Cip1) regulation by transforming growth factor-beta. J Biol Chem 2000; 275: 29244–29256.

    Article  CAS  Google Scholar 

  35. Mulder KM, Levine AE, Hernandez X, McKnight MK, Brattain DE, Brattain MG . Modulation of c-myc by transforming growth factor-beta in human colon carcinoma cells. Biochem Biophys Res Commun 1988; 150: 711–716.

    Article  CAS  Google Scholar 

  36. Zhang XY, Pfeiffer HK, Mellert HS, Stanek TJ, Sussman RT, Kumari A et al2011. Inhibition of the single downstream target BAG1 activates the latent apoptotic potential of MYC. Mol Cell Biol 2011; 31: 5037–5045.

    Article  CAS  Google Scholar 

  37. Myant K, Sansom OJ . Wnt/Myc interactions in intestinal cancer: partners in crime. Exp Cell Res 2011; 317: 2725–2731.

    Article  CAS  Google Scholar 

  38. Siegel PM, Massagué J . Cytostatic and apoptotic actions of TGF-beta in homeostasis and cancer. Nat Rev Cancer 2003; 3: 807–821.

    Article  CAS  Google Scholar 

  39. Friedman E, Gold LI, Klimstra D, Zeng ZS, Winawer S, Cohen A . High levels of transforming growth factor beta-1 correlate with disease progression in human colon cancer. Cancer Epidemiol Biomarkers Prev 1995; 4: 549–554.

    CAS  PubMed  Google Scholar 

  40. Tsushima H, Kawata S, Tamura S, Ito N, Shirai Y, Kiso S et al. High levels of transforming growth factor beta 1 in patients with colorectal cancer: association with disease progression. Gastroenterology 1996; 110: 375–382.

    Article  CAS  Google Scholar 

  41. Picon A, Gold LI, Wang J, Cohen A, Friedman E . A subset of metastatic human colon cancers expresses elevated levels of transforming growth factor beta1. Cancer. Epidemiol Biomarkers Prev 1998; 7: 497–504.

    CAS  Google Scholar 

  42. Xu WQ, Jiang XC, Zheng L, Yu YY, Tang JM . Expression of TGF-beta1, TbetaRII and Smad4 in colorectal carcinoma. Exp Mol Pathol 2007; 82: 284–291.

    Article  CAS  Google Scholar 

  43. Zhang YE . Non-Smad pathways in TGF-beta signalling. Cell Res 2009; 19: 128–139.

    Article  CAS  Google Scholar 

  44. Hong W, Baniahmad A, Liu Y, Li H . Bag-1M is a component of the in vivo DNA-glucocorticoid receptor complex at hormone-regulated promoter. J Mol Biol 2008; 384: 22–30.

    Article  CAS  Google Scholar 

  45. Hong W, Baniahmad A, Li J, Chang C, Gao W, Liu Y . Bag-1M inhibits the transactivation of the glucocorticoid receptor via recruitment of corepressors. FEBS Lett 2009; 583: 2451–2456.

    Article  CAS  Google Scholar 

  46. Lee KY, Ito K, Hayashi R, Jazrawi EP, Barnes PJ, Adcock IM . NF-kappaB and activator protein 1 response elements and the role of histone modifications in IL-1beta-induced TGF-beta1 gene transcription. J Immunol 2006; 176: 603–615.

    Article  CAS  Google Scholar 

  47. Sharp A, Cutress RI, Johnson PW, Packham G, Townsend PA . Short peptides derived from the BAG-1 C-terminus inhibit the interaction between BAG-1 and HSC70 and decrease breast cancer cell growth. FEBS Lett 2009; 583: 3405–3411.

    Article  CAS  Google Scholar 

  48. Sharp A, Crabb SJ, Johnson PW, Hague A, Cutress R, Townsend PA et al. Thioflavin S (NSC71948) interferes with Bcl-2-associated athanogene (BAG-1)-mediated protein-protein interactions. J Pharmacol Exp Ther 2009; 331: 680–689.

    Article  CAS  Google Scholar 

  49. Hanahan D, Weinberg RA . Hallmarks of cancer: the next generation. Cell 2011; 144: 646–674.

    Article  CAS  Google Scholar 

  50. Paraskeva C, Finerty S, Mountford RA, Powell SC . Specific cytogenetic abnormalities in two new human colorectal adenomaderived epithelial cell lines. Cancer Res 1989; 49: 1282–1286.

    CAS  Google Scholar 

  51. Williams AC, Browne SJ, Yeudal WA, Paterson IC, Marshall CJ, Lane DP et al. Molecular events including p53 and k-ras alterations in the in vitro progression of a human colorectal adenoma cell line to an adenocarcinoma. Oncogene 1993; 8: 3063–3072.

    CAS  Google Scholar 

  52. Strathdee CA, McLeod MR, Hall JR . Efficient control of tetracycline-responsive gene expression from an autoregulated bi-directional expression vector. Gene 1999; 229: 21–29.

    Article  CAS  Google Scholar 

  53. Brimmell M, Burns JS, Munson P, McDonald L, O’Hare MJ, Lakhani SR et al. High level expression of differentially localized BAG-1 isoforms in some oestrogen receptor-positive human breast cancers. Br J Cancer 1999; 81: 1042–1051.

    Article  CAS  Google Scholar 

  54. Kaidi A, Williams AC, Paraskeva C . Interaction between beta-catenin and HIF-1 promotes cellular adaptation to hypoxia. Nat Cell Biol 2007; 9: 210–217.

    Article  CAS  Google Scholar 

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Acknowledgements

This work was funded by a Cancer Research UK programme grant, the Citrina Foundation, CORE and by the John James Bristol Foundation. We thank members of the CP group for useful discussion.

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Authors and Affiliations

  1. School of Cellular and Molecular Medicine, University of Bristol, Bristol, UK

    V R Skeen, T J Collard, S L Southern, A Greenhough, C Paraskeva & A C Williams

  2. School of Oral and Dental Sciences, University of Bristol, Bristol, UK

    A Hague

  3. Cancer Sciences, School of Medicine, University of Southampton, Southampton General Hospital, Southampton, UK

    P A Townsend

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Skeen, V., Collard, T., Southern, S. et al. BAG-1 suppresses expression of the key regulatory cytokine transforming growth factor β (TGF-β1) in colorectal tumour cells. Oncogene 32, 4490–4499 (2013). https://doi.org/10.1038/onc.2012.480

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