Skip to main content

Advertisement

SpringerLink
Log in

Downregulation of reversion-inducing-cysteine-rich protein with Kazal motifs (RECK) is associated with enhanced expression of matrix metalloproteinases and cholangiocarcinoma metastases

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

Abstract

Background

Reversion-inducing-cysteine-rich protein with Kazal motifs (RECK) has been implicated in the attenuation of tumor metastasis by negatively regulating metalloproteinase (MMP) levels. RECK gene expression is downregulated in many solid tumors, with this downregulation being associated with poor prognosis. This study evaluated the role of RECK in cholangiocarcinoma (CCA).

Methods

The expression of RECK, MMP-2, and MMP-9 in paraffin sections of hamster and human CCA specimens was analyzed by immunohistochemistry. Functional analysis of RECK was performed in RECK small interfering (si) RNA knockdown CCA cell lines. The effect of aspirin on RECK status and function was evaluated using Western blotting, gelatin zymography, invasion and proliferation assays, and PhosphoELISArray analysis of Ras downstream mediators.

Results

Hamster tissues showed high RECK expression in hyperplastic biliary duct epithelia, low RECK expression in precancerous lesions, and no RECK expression in CCA. In human specimens, RECK was highly expressed in normal biliary cells, whereas intrahepatic CCA showed low levels of expression. Downregulation of RECK was correlated with tumor metastasis (P < 0.01) and shorter patient survival (P < 0.02). RECK expression levels were inversely correlated with MMP-2 and MMP-9 expression (P < 0.05). SiRNA RECK-depleted M139 CCA cells exhibited increased MMP-2/-9 gelatinase activities and invasiveness. Aspirin (500 μM) demonstrated myriad effects in human CCA cell lines, including growth suppression, reduced phosphorylation of Akt/Erk/c-Jun, elevation of RECK expression, inhibition of MMP-2/MMP-9 activity, and enhanced invasiveness.

Conclusions

RECK functions as a metastasis suppressor in CCA; upregulation of RECK expression could provide a potential therapy to improve the prognosis of this type of cancer.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. McLean L, Patel T. Racial and ethnic variations in the epidemiology of intrahepatic cholangiocarcinoma in the United States. Liver Int. 2006;26(9):1047–53.

    Article  PubMed  Google Scholar 

  2. Haswell-Elkins MR, Mairiang E, Mairiang P, Chaiyakum J, Chamadol N, Loapaiboon V, et al. Cross-sectional study of Opisthorchis viverrini infection and cholangiocarcinoma in communities within a high-risk area in northeast Thailand. Int J Cancer. 1994;59(4):505–9.

    Article  PubMed  CAS  Google Scholar 

  3. Sripa B, Kaewkes S, Sithithaworn P, Mairiang E, Laha T, Smout M, et al. Liver fluke induces cholangiocarcinoma. PLoS Med. 2007;4(7):e201.

    Article  PubMed  Google Scholar 

  4. Takahashi C, Sheng Z, Horan TP, Kitayama H, Maki M, Hitomi K, et al. Regulation of matrix metalloproteinase-9 and inhibition of tumor invasion by the membrane-anchored glycoprotein RECK. Proc Natl Acad Sci USA. 1998;95(22):13221–6.

    Article  PubMed  CAS  Google Scholar 

  5. Oh J, Takahashi R, Kondo S, Mizoguchi A, Adachi E, Sasahara RM, et al. The membrane-anchored MMP inhibitor RECK is a key regulator of extracellular matrix integrity and angiogenesis. Cell. 2001;107(6):789–800.

    Article  PubMed  CAS  Google Scholar 

  6. Masamune A, Shimosegawa T. Signal transduction in pancreatic stellate cells. J Gastroenterol. 2009;44(4):249–60.

    Article  PubMed  Google Scholar 

  7. Clark JC, Thomas DM, Choong PF, Dass CR. RECK—a newly discovered inhibitor of metastasis with prognostic significance in multiple forms of cancer. Cancer Metastasis Rev. 2007;26(3–4):675–83.

    Article  PubMed  CAS  Google Scholar 

  8. Furumoto K, Arii S, Mori A, Furuyama H, Gorrin Rivas MJ, Nakao T, et al. RECK gene expression in hepatocellular carcinoma: correlation with invasion-related clinicopathological factors and its clinical significance. Reverse-inducing-cysteine-rich protein with Kazal motifs. Hepatology. 2001;33(1):189–95.

    Article  PubMed  CAS  Google Scholar 

  9. Masui T, Doi R, Koshiba T, Fujimoto K, Tsuji S, Nakajima S, et al. RECK expression in pancreatic cancer: its correlation with lower invasiveness and better prognosis. Clin Cancer Res. 2003;9(5):1779–84.

    PubMed  CAS  Google Scholar 

  10. Takenaka K, Ishikawa S, Kawano Y, Yanagihara K, Miyahara R, Otake Y, et al. Expression of a novel matrix metalloproteinase regulator, RECK, and its clinical significance in resected non-small cell lung cancer. Eur J Cancer. 2004;40(10):1617–23.

    Article  PubMed  CAS  Google Scholar 

  11. Takenaka K, Ishikawa S, Yanagihara K, Miyahara R, Hasegawa S, Otake Y, et al. Prognostic significance of reversion-inducing cysteine-rich protein with Kazal motifs expression in resected pathologic stage IIIA N2 non-small-cell lung cancer. Ann Surg Oncol. 2005;12(10):817–24.

    Article  PubMed  Google Scholar 

  12. Song SY, Son HJ, Nam E, Rhee JC, Park C. Expression of reversion-inducing-cysteine-rich protein with Kazal motifs (RECK) as a prognostic indicator in gastric cancer. Eur J Cancer. 2006;42(1):101–8.

    Article  PubMed  CAS  Google Scholar 

  13. van der Jagt MF, Sweep FC, Waas ET, Hendriks T, Ruers TJ, Merry AH, et al. Correlation of reversion-inducing cysteine-rich protein with Kazal motifs (RECK) and extracellular matrix metalloproteinase inducer (EMMPRIN), with MMP-2, MMP-9, and survival in colorectal cancer. Cancer Lett. 2006;237(2):289–97.

    Article  PubMed  Google Scholar 

  14. Rabien A, Burkhardt M, Jung M, Fritzsche F, Ringsdorf M, Schicktanz H, et al. Decreased RECK expression indicating proteolytic imbalance in prostate cancer is associated with higher tumor aggressiveness and risk of prostate-specific antigen relapse after radical prostatectomy. Eur Urol. 2007;51(5):1259–66.

    Article  PubMed  CAS  Google Scholar 

  15. Nagini S, Letchoumy PV, Thangavelu A, Ramachandran CR. Of humans and hamsters: a comparative evaluation of carcinogen activation, DNA damage, cell proliferation, apoptosis, invasion, and angiogenesis in oral cancer patients and hamster buccal pouch carcinomas. Oral Oncol. 2009;45(6):e31–7.

    Article  PubMed  CAS  Google Scholar 

  16. Li Y, Zhang Y, Zheng Q. Expression of RECK gene and MMP-9 in hilar cholangiocarcinoma and its clinical significance. J Huazhong Univ Sci Technol Med Sci. 2005;25(5):552–4.

    Article  PubMed  CAS  Google Scholar 

  17. Takagi S, Simizu S, Osada H. RECK negatively regulates matrix metalloproteinase-9 transcription. Cancer Res. 2009;69(4):1502–8.

    Article  PubMed  CAS  Google Scholar 

  18. Sasahara RM, Takahashi C, Noda M. Involvement of the Sp1 site in ras-mediated downregulation of the RECK metastasis suppressor gene. Biochem Biophys Res Commun. 1999;264(3):668–75.

    Article  PubMed  CAS  Google Scholar 

  19. Liu LT, Chang HC, Chiang LC, Hung WC. Induction of RECK by nonsteroidal anti-inflammatory drugs in lung cancer cells. Oncogene. 2002;21(54):8347–50.

    Article  PubMed  CAS  Google Scholar 

  20. Liu LT, Chang HC, Chiang LC, Hung WC. Histone deacetylase inhibitor up-regulates RECK to inhibit MMP-2 activation and cancer cell invasion. Cancer Res. 2003;63(12):3069–72.

    PubMed  CAS  Google Scholar 

  21. Loilome W, Yongvanit P, Wongkham C, Tepsiri N, Sripa B, Sithithaworn P, et al. Altered gene expression in Opisthorchis viverrini-associated cholangiocarcinoma in hamster model. Mol Carcinog. 2006;45(5):279–87.

    Article  PubMed  CAS  Google Scholar 

  22. Maruyama M, Kobayashi N, Westerman KA, Sakaguchi M, Allain JE, Totsugawa T, et al. Establishment of a highly differentiated immortalized human cholangiocyte cell line with SV40T and hTERT. Transplantation. 2004;77(3):446–51.

    Article  PubMed  CAS  Google Scholar 

  23. Schmittgen TD, Jiang J, Liu Q, Yang L. A high-throughput method to monitor the expression of microRNA precursors. Nucleic Acids Res. 2004;32(4):e43.

    Article  PubMed  Google Scholar 

  24. Namwat N, Amimanan P, Loilome W, Jearanaikoon P, Sripa B, Bhudhisawasdi V, et al. Characterization of 5-fluorouracil-resistant cholangiocarcinoma cell lines. Chemotherapy. 2008;54(5):343–51.

    Article  PubMed  CAS  Google Scholar 

  25. Deryugina EI, Quigley JP. Matrix metalloproteinases and tumor metastasis. Cancer Metastasis Rev. 2006;25(1):9–34.

    Article  PubMed  CAS  Google Scholar 

  26. Shirabe K, Shimada M, Kajiyama K, Hasegawa H, Gion T, Ikeda Y, et al. Expression of matrix metalloproteinase-9 in surgically resected intrahepatic cholangiocarcinoma. Surgery. 1999;126(5):842–6.

    Article  PubMed  CAS  Google Scholar 

  27. Prakobwong S, Pinlaor S, Yongvanit P, Sithithaworn P, Pairojkul C, Hiraku Y. Time profiles of the expression of metalloproteinases, tissue inhibitors of metalloproteases, cytokines and collagens in hamsters infected with Opisthorchis viverrini with special reference to peribiliary fibrosis and liver injury. Int J Parasitol. 2009;39(7):825–35.

    Article  PubMed  CAS  Google Scholar 

  28. Cho CY, Wang JH, Chang HC, Chang CK, Hung WC. Epigenetic inactivation of the metastasis suppressor RECK enhances invasion of human colon cancer cells. J Cell Physiol. 2007;213(1):65–9.

    Article  PubMed  CAS  Google Scholar 

  29. Chang HC, Liu LT, Hung WC. Involvement of histone deacetylation in ras-induced down-regulation of the metastasis suppressor RECK. Cell Signal. 2004;16(6):675–9.

    Article  PubMed  CAS  Google Scholar 

  30. Hsu MC, Chang HC, Hung WC. HER-2/neu represses the metastasis suppressor RECK via ERK and Sp transcription factors to promote cell invasion. J Biol Chem. 2006;281(8):4718–25.

    Article  PubMed  CAS  Google Scholar 

  31. Loayza-Puch F, Yoshida Y, Matsuzaki T, Takahashi C, Kitayama H, Noda M. Hypoxia and RAS-signaling pathways converge on, and cooperatively downregulate, the RECK tumor-suppressor protein through microRNAs. Oncogene. 2010;29(18):2638–48.

    Article  PubMed  CAS  Google Scholar 

  32. Xiang S, Sun Z, He Q, Yan F, Wang Y, Zhang J. Aspirin inhibits ErbB2 to induce apoptosis in cervical cancer cells. Med Oncol. 2010;27(2):379–87.

    Article  PubMed  CAS  Google Scholar 

  33. Ko HW, Park KY, Kim H, Han PL, Kim YU, Gwag BJ, et al. Ca2+-mediated activation of c-Jun N-terminal kinase and nuclear factor kappa B by NMDA in cortical cell cultures. J Neurochem. 1998;71(4):1390–5.

    Article  PubMed  CAS  Google Scholar 

  34. Jiao X, Katiyar S, Willmarth NE, Liu M, Ma X, Flomenberg N, et al. c-Jun induces mammary epithelial cellular invasion and breast cancer stem cell expansion. J Biol Chem. 2010;285(11):8218–26.

    Article  PubMed  CAS  Google Scholar 

  35. Jacobs EJ, Thun MJ, Bain EB, Rodriguez C, Henley SJ, Calle EE. A large cohort study of long-term daily use of adult-strength aspirin and cancer incidence. J Natl Cancer Inst. 2007;99(8):608–15.

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We would like to thank the Liver Fluke and Cholangiocarcinoma Research Center for its support by a grant fund for the MS.c program, to J.P. This project was co-supported by a Mid-Career Grant (RSA5280007), Thailand Research Fund, to N.N.; and by an Invitation Research grant by the Faculty of Medicine (I52111) to J.P., Khon Kaen University, Thailand. We thank Professor Naoya Kobayashi (Okayama University, Japan), who kindly supplied the human MMNK1 cell line; and we thank the National Cancer Center of Singapore for kindly supplying the HucCT1 and EGI-1 cell lines used for this study. The content of this manuscript was edited and commented upon by Professor Prapon Wilairat, Mahidol University, Thailand. The English editing of this manuscript was kindly performed by Dr. Rahul Kuver, University of Washington, Seattle, WA, USA.

Author information

Authors and Affiliations

  1. Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand

    N. Namwat, J. Puetkasichonpasutha, W. Loilome, P. Yongvanit, A. Techasen & S. Wongkham

  2. Department of Pathology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand

    A. Puapairoj & B. Sripa

  3. Department of Pharmacology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand

    W. Tassaneeyakul

  4. Department of Surgery, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand

    N. Khuntikeo

  5. Liver Fluke and Cholangiocarcinoma Research Center, Khon Kaen University, Khon Kaen, Thailand

    N. Namwat, J. Puetkasichonpasutha, W. Loilome, P. Yongvanit, A. Techasen, A. Puapairoj, B. Sripa, W. Tassaneeyakul, N. Khuntikeo & S. Wongkham

Authors
  1. N. Namwat
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Puetkasichonpasutha
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. W. Loilome
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. P. Yongvanit
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A. Techasen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. A. Puapairoj
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. B. Sripa
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. W. Tassaneeyakul
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. N. Khuntikeo
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. S. Wongkham
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to N. Namwat.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Namwat, N., Puetkasichonpasutha, J., Loilome, W. 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 46, 664–675 (2011). https://doi.org/10.1007/s00535-010-0345-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00535-010-0345-y

Keywords

Search

Navigation