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WTAP regulates migration and invasion of cholangiocarcinoma cells

  • Original Article—Liver, Pancreas, and Biliary Tract
  • Published:
Journal of Gastroenterology Aims and scope Submit manuscript

Abstract

Background

Wilms’ tumor 1-associating protein (WTAP) is a nuclear protein that has been associated with the regulation of proliferation and apoptosis. Although its dynamic expression and physiological functions in vascular cells have been reported, its expression and roles in cholangiocarcinoma cells are poorly characterized.

Methods

To examine the expression of WTAP in patient tissues, we performed immunohistochemistry. To examine motility of cholangiocarcinoma cells, we employed Boyden chamber, wound healing and Matrigel invasion assays, and a liver xenograft model.

Results

Immunohistochemistry in patient tissues showed WTAP overexpression in cholangiocarcinoma tissues and correlation of WTAP expression with metastasis of cholangiocarcinoma cells. Overexpression or knockdown of WTAP significantly increased or decreased the motility of cholangiocarcinoma cells. Moreover, WTAP overexpression or knockdown significantly increased or decreased tumorigenicity of cholangiocarcinoma cells in an orthotopic xenograft model. Furthermore, microarray study showed that WTAP induce the expressions of MMP7, MMP28, cathepsin H and Muc1.

Conclusion

WTAP is overexpressed in cholangiocarcinoma and regulates motility of cholangiocarcinoma cells

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References

  1. Kato I, Kuroishi T, Tominaga S. Descriptive epidemiology of subsites of cancers of the liver, biliary tract and pancreas in Japan. Jpn J Clin Oncol. 1990;20:232–7.

    CAS  PubMed  Google Scholar 

  2. Taylor-Robinson SD, Foster GR, Arora S, Hargreaves S, Thomas HC. Increase in primary liver cancer in the UK, 1979–94. Lancet. 1997;350:1142–3.

    Article  CAS  PubMed  Google Scholar 

  3. Olnes MJ, Erlich R. A review and update on cholangiocarcinoma. Oncology. 2004;66:167–79.

    Article  PubMed  Google Scholar 

  4. Shaib YH, Davila JA, McGlynn K, El-Serag HB. Rising incidence of intrahepatic cholangiocarcinoma in the United States: a true increase? J Hepatol. 2004;40:472–7.

    Article  PubMed  Google Scholar 

  5. Little NA, Hastie ND, Davies RC. Identification of WTAP, a novel Wilms’ tumour 1-associating protein. Hum Mol Genet. 2000;9:2231–9.

    Article  CAS  PubMed  Google Scholar 

  6. Horiuchi K, Umetani M, Minami T, Okayama H, Takada S, Yamamoto M, et al. Wilms’ tumor 1-associating protein regulates G2/M transition through stabilization of cyclin A2 mRNA. Proc Natl Acad Sci USA. 2006;103:17278–83.

    Article  CAS  PubMed  Google Scholar 

  7. Naruse C, Fukusumi Y, Kakiuchi D, Asano M. A novel gene trapping for identifying genes expressed under the control of specific transcription factors. Biochem Biophys Res Commun. 2007;361:109–15.

    Article  CAS  PubMed  Google Scholar 

  8. Small TW, Bolender Z, Bueno C, O’Neil C, Nong Z, Rushlow W, et al. Wilms’ tumor 1-associating protein regulates the proliferation of vascular smooth muscle cells. Circ Res. 2006;99:1338–46.

    Article  CAS  PubMed  Google Scholar 

  9. Small TW, Penalva LO, Pickering JG. Vascular biology and the sex of flies: regulation of vascular smooth muscle cell proliferation by wilms’ tumor 1-associating protein. Trends Cardiovasc Med. 2007;17:230–4.

    Article  CAS  PubMed  Google Scholar 

  10. Small TW, Pickering JG. Nuclear degradation of Wilms tumor 1-associating protein and survivin splice variant switching underlie IGF-1-mediated survival. J Biol Chem. 2009;284:24684–95.

    Article  CAS  PubMed  Google Scholar 

  11. Lee SB, Huang K, Palmer R, Truong VB, Herzlinger D, Kolquist KA, et al. The Wilms tumor suppressor WT1 encodes a transcriptional activator of amphiregulin. Cell. 1999;98:663–73.

    Article  CAS  PubMed  Google Scholar 

  12. Mayo MW, Wang CY, Drouin SS, Madrid LV, Marshall AF, Reed JC, et al. WT1 modulates apoptosis by transcriptionally upregulating the bcl-2 proto-oncogene. EMBO J. 1999;18:3990–4003.

    Article  CAS  PubMed  Google Scholar 

  13. Saitoh N, Spahr CS, Patterson SD, Bubulya P, Neuwald AF, Spector DL. Proteomic analysis of interchromatin granule clusters. Mol Biol Cell. 2004;15:3876–90.

    Article  CAS  PubMed  Google Scholar 

  14. Zhou Z, Licklider LJ, Gygi SP, Reed R. Comprehensive proteomic analysis of the human spliceosome. Nature. 2002;419:182–5.

    Article  CAS  PubMed  Google Scholar 

  15. Ortega A, Niksic M, Bachi A, Wilm M, Sanchez L, Hastie N, et al. Biochemical function of female-lethal (2)D/Wilms’ tumor suppressor-1-associated proteins in alternative pre-mRNA splicing. J Biol Chem. 2003;278:3040–7.

    Article  CAS  PubMed  Google Scholar 

  16. Jeon TY, Han ME, Lee YW, Lee YS, Kim GH, Song GA, et al. Overexpression of stathmin1 in the diffuse type of gastric cancer and its roles in proliferation and migration of gastric cancer cells. Br J Cancer. 2010;102:710–8.

    Article  CAS  PubMed  Google Scholar 

  17. Hisatsune A, Nakayama H, Kawasaki M, Horie I, Miyata T, Isohama Y, et al. Anti-MUC1 antibody inhibits EGF receptor signaling in cancer cells. Biochem Biophys Res Commun. 2011;405:377–81.

    Article  CAS  PubMed  Google Scholar 

  18. Itatsu K, Sasaki M, Yamaguchi J, Ohira S, Ishikawa A, Ikeda H, et al. Cyclooxygenase-2 is involved in the up-regulation of matrix metalloproteinase-9 in cholangiocarcinoma induced by tumor necrosis factor-alpha. Am J Pathol. 2009;174:829–41.

    Article  CAS  PubMed  Google Scholar 

  19. Ohira S, Sasaki M, Harada K, Sato Y, Zen Y, Isse K, et al. Possible regulation of migration of intrahepatic cholangiocarcinoma cells by interaction of CXCR4 expressed in carcinoma cells with tumor necrosis factor-alpha and stromal-derived factor-1 released in stroma. Am J Pathol. 2006;168:1155–68.

    Article  CAS  PubMed  Google Scholar 

  20. Zhang K, Zhaos J, Liu X, Yan B, Chen D, Gao Y, et al. Activation of NF-B upregulates Snail and consequent repression of E-cadherin in cholangiocarcinoma cell invasion. Hepatogastroenterology. 2011;58:1–7.

    Article  CAS  PubMed  Google Scholar 

  21. Ishimura N, Isomoto H, Bronk SF, Gores GJ. Trail induces cell migration and invasion in apoptosis-resistant cholangiocarcinoma cells. Am J Physiol Gastrointest Liver Physiol. 2006;290:G129–36.

    Article  CAS  PubMed  Google Scholar 

  22. Lu D, Han C, Wu T. Microsomal prostaglandin E synthase-1 inhibits PTEN and promotes experimental cholangiocarcinogenesis and tumor progression. Gastroenterology. 2011;140:2084–94.

    Article  CAS  PubMed  Google Scholar 

  23. Leelawat K, Leelawat S, Tepaksorn P, Rattanasinganchan P, Leungchaweng A, Tohtong R, et al. Involvement of c-Met/hepatocyte growth factor pathway in cholangiocarcinoma cell invasion and its therapeutic inhibition with small interfering RNA specific for c-Met. J Surg Res. 2006;136:78–84.

    Article  CAS  PubMed  Google Scholar 

  24. Lee MJ, Yu GR, Yoo HJ, Kim JH, Yoon BI, Choi YK, et al. ANXA8 down-regulation by EGF-FOXO4 signaling is involved in cell scattering and tumor metastasis of cholangiocarcinoma. Gastroenterology. 2009;137:1138–50, 50 e1–9.

    Google Scholar 

  25. Chen TC, Jan YY, Yeh TS. K-ras mutation is strongly associated with perineural invasion and represents an independent prognostic factor of intrahepatic cholangiocarcinoma after hepatectomy. Ann Surg Oncol. 2012;19(Suppl 3):S675–81.

    Article  PubMed  Google Scholar 

  26. Ukita Y, Kato M, Terada T. Gene amplification and mRNA and protein overexpression of c-erbB-2 (HER-2/neu) in human intrahepatic cholangiocarcinoma as detected by fluorescence in situ hybridization, in situ hybridization, and immunohistochemistry. J Hepatol. 2002;36:780–5.

    Article  CAS  PubMed  Google Scholar 

  27. Benckert C, Jonas S, Cramer T, Von Marschall Z, Schafer G, Peters M, et al. Transforming growth factor beta 1 stimulates vascular endothelial growth factor gene transcription in human cholangiocellular carcinoma cells. Cancer Res. 2003;63:1083–92.

    CAS  PubMed  Google Scholar 

  28. Araki K, Shimura T, Suzuki H, Tsutsumi S, Wada W, Yajima T, et al. E/N-cadherin switch mediates cancer progression via TGF-beta-induced epithelial-to-mesenchymal transition in extrahepatic cholangiocarcinoma. Br J Cancer. 2011;105:1885–93.

    Article  CAS  PubMed  Google Scholar 

  29. Fukase K, Ohtsuka H, Onogawa T, Oshio H, Ii T, Mutoh M, et al. Bile acids repress E-cadherin through the induction of Snail and increase cancer invasiveness in human hepatobiliary carcinoma. Cancer Sci. 2008;99:1785–92.

    Article  CAS  PubMed  Google Scholar 

  30. Oishi N, Kumar MR, Roessler S, Ji J, Forgues M, Budhu A, et al. Transcriptomic profiling reveals hepatic stem-like gene signatures and interplay of miR-200c and epithelial-mesenchymal transition in intrahepatic cholangiocarcinoma. Hepatology. 2012;56:1792–803.

    Article  CAS  PubMed  Google Scholar 

  31. Shi RY, Yang XR, Shen QJ, Yang LX, Xu Y, Qiu SJ, et al. High expression of Dickkopf-related protein 1 is related to lymphatic metastasis and indicates poor prognosis in intrahepatic cholangiocarcinoma patients after surgery. Cancer.

  32. Sugimachi K, Taguchi K, Aishima S, Tanaka S, Shimada M, Kajiyama K, et al. Altered expression of beta-catenin without genetic mutation in intrahepatic cholangiocarcinoma. Mod Pathol. 2001;14:900–5.

    Article  CAS  PubMed  Google Scholar 

  33. Lee S, Kim WH, Jung HY, Yang MH, Kang GH. Aberrant CpG island methylation of multiple genes in intrahepatic cholangiocarcinoma. Am J Pathol. 2002;161:1015–22.

    Article  CAS  PubMed  Google Scholar 

  34. Kim KS, Jin DI, Yoon S, Baek SY, Kim BS, Oh SO. Expression and roles of NUPR1 in cholangiocarcinoma cells. Anat Cell Biol. 2012;45:17–25.

    Article  PubMed  Google Scholar 

  35. Baek S, Lee YW, Yoon S, Baek SY, Kim BS, Oh SO. CDH3/P-Cadherin regulates migration of HuCCT1 cholangiocarcinoma cells. Anat Cell Biol. 2010;43:110–7.

    Article  PubMed  Google Scholar 

  36. Keeratichamroen S, Leelawat K, Thongtawee T, Narong S, Aegem U, Tujinda S, et al. Expression of CD24 in cholangiocarcinoma cells is associated with disease progression and reduced patient survival. Int J Oncol. 2011;39:873–81.

    CAS  PubMed  Google Scholar 

  37. Pongcharoen P, Jinawath A, Tohtong R. Silencing of CD44 by siRNA suppressed invasion, migration and adhesion to matrix, but not secretion of MMPs, of cholangiocarcinoma cells. Clin Exp Metastasis. 2011;28:827–39.

    Article  CAS  PubMed  Google Scholar 

  38. Junking M, Wongkham C, Sripa B, Sawanyawisuth K, Araki N, Wongkham S. Decreased expression of galectin-3 is associated with metastatic potential of liver fluke-associated cholangiocarcinoma. Eur J Cancer. 2008;44:619–26.

    Article  CAS  PubMed  Google Scholar 

  39. Techasen A, Loilome W, Namwat N, Takahashi E, Sugihara E, Puapairoj A, et al. Myristoylated alanine-rich C kinase substrate phosphorylation promotes cholangiocarcinoma cell migration and metastasis via the protein kinase C-dependent pathway. Cancer Sci. 2010;101:658–65.

    Article  CAS  PubMed  Google Scholar 

  40. Nuntagowat C, Leelawat K, Tohtong R. NGAL knockdown by siRNA in human cholangiocarcinoma cells suppressed invasion by reducing NGAL/MMP-9 complex formation. Clin Exp Metastasis. 2010;27:295–305.

    Article  CAS  PubMed  Google Scholar 

  41. Min JK, Kim JM, Li S, Lee JW, Yoon H, Ryu CJ, et al. L1 cell adhesion molecule is a novel therapeutic target in intrahepatic cholangiocarcinoma. Clin Cancer Res. 2010;16:3571–80.

    Article  CAS  PubMed  Google Scholar 

  42. Jeong CY, Hah YS, Cho BI, Lee SM, Joo YT, Jung EJ, et al. Fatty acid-binding protein 5 promotes cell proliferation and invasion in human intrahepatic cholangiocarcinoma. Oncol Rep. 2012;28:1283–92.

    CAS  PubMed  Google Scholar 

  43. Huang Q, Liu C, Wang C, Hu Y, Qiu L, Xu P. Neurotransmitter gamma-aminobutyric acid-mediated inhibition of the invasive ability of cholangiocarcinoma cells. Oncol Lett. 2011;2:519–23.

    CAS  PubMed  Google Scholar 

  44. Namwat N, Puetkasichonpasutha J, Loilome W, Yongvanit P, Techasen A, Puapairoj A, et al. Downregulation of reversion-inducing-cysteine-rich protein with Kazal motifs (RECK) is associated with enhanced expression of matrix metalloproteinases and cholangiocarcinoma metastases. J Gastroenterol. 2011;46:664–75.

    Article  CAS  PubMed  Google Scholar 

  45. Zeng B, Li Z, Chen R, Guo N, Zhou J, Zhou Q, et al. Epigenetic regulation of miR-124 by Hepatitis C Virus core protein promotes migration and invasion of intrahepatic cholangiocarcinoma cells by targeting SMYD3. FEBS Lett. 2011;586:3271–8.

    Article  CAS  Google Scholar 

  46. Xiong GP, Zhang JX, Gu SP, Wu YB, Liu JF. Overexpression of ECM1 contributes to migration and invasion in cholangiocarcinoma cell. Neoplasma. 2012;59:409–15.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported by Medical Research Institute Grant (2009-22), Pusan National University Hospital, the medical research centre program of Ministry of Education, Science and Technology/Korea Science and Engineering Foundation (2011-0006190), a grant from the National R&D Program for Cancer Control, Ministry for Health, Welfare and Family Affairs, Republic of Korea (0920050). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The biospecimens for this study were provided by the Pusan National University Hospital, a member of the National Biobank of Korea, which is supported by the Ministry of Health, Welfare and Family Affairs. All samples derived from the National Biobank of Korea were obtained with informed consent under institutional review board-approved protocols.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Departments of Surgery, School of Medicine, Pusan National University, Busan, Republic of Korea

    Hong-Jae Jo

  2. Departments of Anatomy, School of Medicine, Pusan National University, Beomeo-ri, Mulgeum-eup, Yangsan-si, Gyeongsangnam-do, 626-870, Republic of Korea

    Hye-Eun Shim, Myoung-Eun Han, Hyun-Jung Kim, Ki-Sun Kim, Sungmin Baek & Sae-Ock Oh

  3. Medical Research Center for Ischemic Tissue Regeneration, Pusan National University, Beomeo-ri, Mulgeum-eup, Yangsan-si, Gyeongsangnam-do, 626-870, Republic of Korea

    Hye-Eun Shim, Myoung-Eun Han, Hyun-Jung Kim, Sungmin Baek & Sae-Ock Oh

  4. Departments of Pathology, School of Medicine, Pusan National University, Beomeo-ri, Mulgeum-eup, Yangsan-si, Gyeongsangnam-do, 626-870, Republic of Korea

    Kyung-Un Choi

  5. Departments of Forensic Medicine, School of Medicine, Pusan National University, Beomeo-ri, Mulgeum-eup, Yangsan-si, Gyeongsangnam-do, 626-870, Republic of Korea

    Gi-Yeong Hur

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  1. Hong-Jae Jo
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Corresponding author

Correspondence to Sae-Ock Oh.

Additional information

H.-J. Jo and H.-E. Shim equally contributed to this work.

Electronic supplementary material

535_2013_748_MOESM1_ESM.pdf

Supplementary Fig. 1. Staining patterns of WTAP in hepatocytes and epithelial cells of the small intestine. Although not all hepatocytes expressed WTAP, its staining pattern in hepatocytes was mostly cytoplasmic or membranous. However the nuclear staining was clearly observed in epithelial cells of the small intestine. Scale bar, 50 μm (PDF 212 kb)

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Jo, HJ., Shim, HE., Han, ME. et al. WTAP regulates migration and invasion of cholangiocarcinoma cells. J Gastroenterol 48, 1271–1282 (2013). https://doi.org/10.1007/s00535-013-0748-7

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  • DOI: https://doi.org/10.1007/s00535-013-0748-7

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