Tumor Biology

, Volume 37, Issue 7, pp 9399–9410 | Cite as

Low expression of fibulin-1 correlates with unfavorable prognosis in gastric cancer

  • Liang Feng
  • Chan Yao
  • Peng Li
  • Ying Feng
  • Fei Wang
  • Yi-Fei Liu
  • Yi-Bing Guo
  • Qin-Sheng Mao
  • Wan-Jiang Xue
Original Article

Abstract

The tumor-suppressing role of fibulin-1 has been described in several types of cancers. However, the expression and role of fibulin-1 in the development and progression of gastric cancer (GC) remain largely unknown. In this study, RT-PCR and immunochemistry were used to detect the fibulin-1 expression in GC samples. We have found that the fibulin-1 protein and mRNA levels were downregulated in GC. When investigating the correlation between fibulin-1 expression and clinicopathological characteristics, we have found that low fibulin-1 protein expression was associated with poor tumor differentiation and advanced N stage. Low fibulin-1 protein expression was also an independent prognostic factor for patient survival. To clarify the reason of fibulin-1 downregulation in GC, the mRNA expression and methylation status of fibulin-1 were examined in GC fresh tissue samples (n = 36). We found that the transcriptional expression of fibulin-1 was negatively associated with fibulin-1 promoter hypermethylation, and fibulin-1 hypermethylation was associated with Helicobacter pylori infection. Finally, the effects of fibulin-1 overexpression on cell proliferation and apoptosis were examined. We have found that fibulin-1 overexpression suppressed the growth of GC both in vitro and in vivo and induced apoptosis by increasing cleaved caspase-3 expression. In conclusion, fibulin-1 acts as a tumor suppressor gene, is frequently hypermethylated in GC, and can potentially serve as a useful biomarker for patient prognosis.

Keywords

Fibulin-1 Gastric carcinogenesis Epigenetic Tumor suppressor gene Prognosis 

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (no. 81000984), the Six Talent Peaks Project of the Jiangsu Province (2011-WS066), and the Social Undertakings of Science and Technology Innovation and Demonstration Project of Nantong City (HS2014042).

Compliance with ethical standards

Conflicts of interest

None

References

  1. 1.
    May M. Statistics: attacking an epidemic. Nature. 2014;509:S50–1.CrossRefPubMedGoogle Scholar
  2. 2.
    Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, et al. Cancer incidence and mortality worldwide: sources, methods and major patterns in globocan 2012. Int J Cancer. 2015;136:E359–86.CrossRefPubMedGoogle Scholar
  3. 3.
    Pourhoseingholi MA, Vahedi M, Baghestani AR. Burden of gastrointestinal cancer in Asia: an overview. Gastroenterol Hepatol Bed Bench. 2015;8:19–27.PubMedPubMedCentralGoogle Scholar
  4. 4.
    De Angelis R, Sant M, Coleman MP, Francisci S, Baili P, Pierannunzio D, et al. Cancer survival in europe 1999–2007 by country and age: results of eurocare-5—a population-based study. Lancet Oncol. 2014;15:23–34.CrossRefPubMedGoogle Scholar
  5. 5.
    Ilson DH. Angiogenesis in gastric cancer: hitting the target? The Lancet. 2014;383:4–6.CrossRefGoogle Scholar
  6. 6.
    Oba K, Paoletti X, Alberts S, Bang YJ, Benedetti J, Bleiberg H, et al. Disease-free survival as a surrogate for overall survival in adjuvant trials of gastric cancer: a meta-analysis. J Natl Cancer Inst. 2013;105:1600–7.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Kusano T, Shiraishi N, Shiroshita H, Etoh T, Inomata M, Kitano S. Poor prognosis of advanced gastric cancer with metastatic suprapancreatic lymph nodes. Ann Surg Oncol. 2013;20:2290–5.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Tsuburaya A, Mizusawa J, Tanaka Y, Fukushima N, Nashimoto A, Sasako M, et al. Neoadjuvant chemotherapy with s-1 and cisplatin followed by d2 gastrectomy with para-aortic lymph node dissection for gastric cancer with extensive lymph node metastasis. Br J Surg. 2014;101:653–60.CrossRefPubMedGoogle Scholar
  9. 9.
    Wadhwa R, Song S, Lee JS, Yao Y, Wei Q, Ajani JA. Gastric cancer-molecular and clinical dimensions. Nat Rev Clin Oncol. 2013;10:643–55.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Gomez-Martin C, Plaza JC, Pazo-Cid R, Salud A, Pons F, Fonseca P, et al. Level of HER2 gene amplification predicts response and overall survival in HER2-positive advanced gastric cancer treated with trastuzumab. J Clin Oncol. 2013;31:4445–52.CrossRefPubMedGoogle Scholar
  11. 11.
    Cancer Genome Atlas Research N. Comprehensive molecular characterization of gastric adenocarcinoma. Nature. 2014;513:202–9.CrossRefGoogle Scholar
  12. 12.
    Brzozowa M, Michalski M, Harabin-Slowinska M, Wojnicz R. The role of tumour microenvironment in gastric cancer angiogenesis. Prz Gastroenterol. 2014;9:325–8.PubMedPubMedCentralGoogle Scholar
  13. 13.
    Bhome R, Bullock MD, Al Saihati HA, Goh RW, Primrose JN, Sayan AE, et al. A top-down view of the tumor microenvironment: structure, cells and signaling. Front Cell Dev Biol. 2015;3:33.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Yamaguchi H, Yoshida N, Takanashi M, Ito Y, Fukami K, Yanagihara K, et al. Stromal fibroblasts mediate extracellular matrix remodeling and invasion of scirrhous gastric carcinoma cells. PLoS One. 2014;9:e85485.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Timpl R, Sasaki T, Kostka G, Chu ML. Fibulins: a versatile family of extracellular matrix proteins. Nat Rev Mol Cell Biol. 2003;4:479–89.CrossRefPubMedGoogle Scholar
  16. 16.
    Chu ML, Tsuda T. Fibulins in development and heritable disease. Birth Defects Res C Embryo Today. 2004;72:25–36.CrossRefPubMedGoogle Scholar
  17. 17.
    Gallagher WM, Currid CA, Whelan LC. Fibulins and cancer: friend or foe? Trends Mol Med. 2005;11:336–40.CrossRefPubMedGoogle Scholar
  18. 18.
    Greene LM, Twal WO, Duffy MJ, McDermott EW, Hill AD, O’Higgins NJ, et al. Elevated expression and altered processing of fibulin-1 protein in human breast cancer. Br J Cancer. 2003;88:871–8.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Wlazlinski A, Engers R, Hoffmann MJ, Hader C, Jung V, Muller M, et al. Downregulation of several fibulin genes in prostate cancer. Prostate. 2007;67:1770–80.CrossRefPubMedGoogle Scholar
  20. 20.
    Kubista B, Klinglmueller F, Bilban M, Pfeiffer M, Lass R, Giurea A, et al. Microarray analysis identifies distinct gene expression profiles associated with histological subtype in human osteosarcoma. Int Orthop. 2011;35:401–11.CrossRefPubMedGoogle Scholar
  21. 21.
    Twal WO, Czirok A, Hegedus B, Knaak C, Chintalapudi MR, Okagawa H, et al. Fibulin-1 suppression of fibronectin-regulated cell adhesion and motility. J Cell Sci. 2001;114:4587–98.PubMedGoogle Scholar
  22. 22.
    Kostka G, Giltay R, Bloch W, Addicks K, Timpl R, Fassler R, et al. Perinatal lethality and endothelial cell abnormalities in several vessel compartments of fibulin-1-deficient mice. Mol Cell Biol. 2001;21:7025–34.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Cheng YY, Jin H, Liu X, Siu JM, Wong YP, Ng EK, et al. Fibulin 1 is downregulated through promoter hypermethylation in gastric cancer. Br J Cancer. 2008;99:2083–7.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Xu YF, Yi Y, Qiu SJ, Gao Q, Li YW, Dai CX, et al. PEBP1 downregulation is associated to poor prognosis in HCC related to hepatitis b infection. J Hepatol. 2010;53:872–9.CrossRefPubMedGoogle Scholar
  25. 25.
    Xue WJ, Li C, Zhou XJ, Guan HG, Qin L, Li P, et al. RASSF1A expression inhibits the growth of hepatocellular carcinoma from Qidong County. J Gastroenterol Hepatol. 2008;23:1448–58.CrossRefPubMedGoogle Scholar
  26. 26.
    Mei H, Lian S, Zhang S, Wang W, Mao Q, Wang H. High expression of ROR2 in cancer cell correlates with unfavorable prognosis in colorectal cancer. Biochem Biophys Res Commun. 2014;453:703–9.CrossRefPubMedGoogle Scholar
  27. 27.
    Feng Y, Xue WJ, Li P, Sha ZY, Huang H, Rui L, et al. RASSF1A hypermethylation is associated with aflatoxin B1 and polycyclic aromatic hydrocarbon exposure in hepatocellular carcinoma. Hepatogastroenterology. 2012;59:1883–8.PubMedGoogle Scholar
  28. 28.
    Hsieh LL, Lin PJ, Chen TC, Ou JT. Frequency of Epstein-Barr virus-associated gastric adenocarcinoma in Taiwan. Cancer Lett. 1998;129:125–9.CrossRefPubMedGoogle Scholar
  29. 29.
    Kanai M, Konda Y, Nakajima T, Izumi Y, Takeuchi T, Chiba T. TGF-alpha inhibits apoptosis of murine gastric pit cells through an NF-kappaB-dependent pathway. Gastroenterology. 2001;121:56–67.CrossRefPubMedGoogle Scholar
  30. 30.
    Otani K, Li X, Arakawa T, Chan FK, Yu J. Epigenetic-mediated tumor suppressor genes as diagnostic or prognostic biomarkers in gastric cancer. Expert Rev Mol Diagn. 2013;13:445–55.CrossRefPubMedGoogle Scholar
  31. 31.
    Chiba T, Marusawa H, Ushijima T. Inflammation-associated cancer development in digestive organs: mechanisms and roles for genetic and epigenetic modulation. Gastroenterology. 2012;143:550–63.CrossRefPubMedGoogle Scholar
  32. 32.
    Xiao W, Wang J, Li H, Xia D, Yu G, Yao W, et al. Fibulin-1 is epigenetically down-regulated and related with bladder cancer recurrence. BMC Cancer. 2014;14:677.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Wu BJ, Zhou ZP, Li WP, Ding W, Wu YZ, Zhou ZW, et al. Abnormal hypermethylation and clinicopathological significance of FBLN1 gene in cutaneous melanoma. Tumour Biol. 2014;35:123–7.CrossRefPubMedGoogle Scholar
  34. 34.
    Kanda M, Nomoto S, Okamura Y, Hayashi M, Hishida M, Fujii T, et al. Promoter hypermethylation of fibulin 1 gene is associated with tumor progression in hepatocellular carcinoma. Mol Carcinog. 2011;50:571–9.CrossRefPubMedGoogle Scholar
  35. 35.
    Feil R, Fraga MF. Epigenetics and the environment: emerging patterns and implications. Nat Rev Genet. 2011;13:97–109.Google Scholar
  36. 36.
    Konturek PC, Konturek SJ, Brzozowski T. Gastric cancer and Helicobacter pylori infection. J Physiol Pharmacol. 2006;57 Suppl 3:51–65.Google Scholar
  37. 37.
    El-Omar EM, Carrington M, Chow WH, McColl KE, Bream JH, Young HA, et al. Interleukin-1 polymorphisms associated with increased risk of gastric cancer. Nature. 2000;404:398–402.CrossRefPubMedGoogle Scholar
  38. 38.
    Hmadcha A, Bedoya FJ, Sobrino F, Pintado E. Methylation-dependent gene silencing induced by interleukin 1beta via nitric oxide production. J Exp Med. 1999;190:1595–604.CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Augusto AC, Miguel F, Mendonca S, Pedrazzoli Jr J, Gurgueira SA. Oxidative stress expression status associated to helicobacter pylori virulence in gastric diseases. Clin Biochem. 2007;40:615–22.CrossRefPubMedGoogle Scholar
  40. 40.
    Argraves WS, Twal WO, Hammad SM, Guffy SL. A novel intracellular fibulin-1d variant binds to the cytoplasmic domain of integrin beta 1 subunit. Matrix Biol. 2015;43:97–108.CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Kobayashi N, Kostka G, Garbe JHO, Keene DR, Bachinger HP, Hanisch FG, et al. A comparative analysis of the fibulin protein family: biochemical characterization, binding interactions, and tissue localization. J Biol Chem. 2007;282:11805–16.CrossRefPubMedGoogle Scholar
  42. 42.
    Qing J, Maher VM, Tran H, Argraves WS, Dunstan RW, McCormick JJ. Suppression of anchorage-independent growth and matrigel invasion and delayed tumor formation by elevated expression of fibulin-1d in human. Oncogene. 1997;15:2159–68.CrossRefPubMedGoogle Scholar
  43. 43.
    Xiao W, Wang J, Li H, Guan W, Xia D, Yu G, et al. Fibulin-1 is down-regulated through promoter hypermethylation and suppresses renal cell carcinoma progression. J Urol. 2013;190:291–301.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Liang Feng
    • 1
  • Chan Yao
    • 2
  • Peng Li
    • 1
  • Ying Feng
    • 1
  • Fei Wang
    • 1
  • Yi-Fei Liu
    • 2
  • Yi-Bing Guo
    • 3
  • Qin-Sheng Mao
    • 1
    • 4
  • Wan-Jiang Xue
    • 1
    • 4
  1. 1.Department of General SurgeryNantong University Affiliated HospitalNantongChina
  2. 2.Department of PathologyNantong University Affiliated HospitalNantongChina
  3. 3.Department of Surgical Comprehensive LaboratoryNantong University Affiliated HospitalNantongChina
  4. 4.Department of Minimally Invasive SurgeryThe Affiliated Hospital of Nantong UniversityNantongChina

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