Tumor Biology

, Volume 36, Issue 2, pp 1081–1089 | Cite as

The tumor-promoting function of ECRG4 in papillary thyroid carcinoma and its related mechanism

Research Article

Abstract

This study aimed to explore the tumor-promoting function of esophageal cancer-related gene 4 (ECRG4) in the papillary thyroid cancer and its related mechanism. ECRG4 Messenger RNA (mRNA) and protein expression analysis in papillary thyroid cancer tissues was performed by quantitative real-time PCR (Q-RT-PCR), Western blot, and immunohistochemistry methods. Ten pairs of fresh samples from the papillary thyroid carcinoma patients were analyzed for ECRG4 promoter CpG island methylation status by bisulfite sequencing analysis. We also transfected ECRG4 into papillary thyroid cancer cell lines W3 and K1 with lentivirus and analyzed ECRG4 functions through evaluating the changes of the proliferation activity, the cell cycle, and the cell apoptosis rate of these transformed cells. We found that ECRG4 expression was upregulated in most papillary thyroid cancer samples (70.0 %, 28 out of 40 papillary thyroid cancer samples) on the protein level, and the ECRG4 mRNA level was also enhanced in tumor tissues compared to their matched nontumor tissues. CpG islands around the ECRG4 promoter region were demethylated in the papillary thyroid cancer samples. At the same time, the upregulated expression of ECRG4 in papillary thyroid cancer cell lines W3 and K1 could promote both the proliferation activity and the cell cycle transition from the G1 phase into the G2 but could not affect the cell apoptosis rate. The expression of ECRG4 is frequently upregulated in a papillary thyroid carcinoma through the demethylation mechanism of CpG islands in the gene promoter region, and the ECRG4 has a tumor-promoting function through inducing the cell cycle transition from the G1 phase to the G2 in papillary thyroid carcinoma cells.

Keywords

ECRG4 Papillary thyroid carcinoma Methylation Tumor-promoting gene 

Notes

Acknowledgments

This study was supported by grants from the Natural Science Foundation of China (81272214 and 81402979), Foundation of Zhejiang Natural Science Foundation (Y2100248), Foundation of Department of Science and Technology of Zhejiang Province (2009C33155 and 2014C33155), Foundation of Zhejiang Health Department (2009A218 and 2014KYA230), Taizhou Science and Technology Bureau (102KY15), and Zhejiang Province Chinese Medicine Study Foundation (2011ZA113).

Conflicts of interest

None

References

  1. 1.
    Mori Y, Ishiguro H, Kuwabara Y, Kimura M, Mitsui A, Kurehara H, et al. Expression of ECRG4 is an independent prognostic factor for poor survival in patients with esophageal squamous cell carcinoma. Oncol Rep. 2007;18:981–5.PubMedGoogle Scholar
  2. 2.
    Li LW, Yu XY, Yang Y, Zhanag CP, Guo LP, Lu SH. Expression of esophageal cancer related gene 4 (ECRG4), a novel tumor suppressor gene, in esophageal cancer and its inhibitory effect on the tumor growth in vitro and in vivo. Int J Cancer. 2009;125:1505–13.CrossRefPubMedGoogle Scholar
  3. 3.
    Matsuzaki J, Torigoe T, Hirohashi Y, Tamura Y, Asanuma H, Nakazawa E, et al. Expression of ECRG4 is associated with lower proliferative potential of esophageal cancer cells. Pathol Int. 2013;63(8):391–7.CrossRefPubMedGoogle Scholar
  4. 4.
    Li LW, Li YY, Li XY, Zhang CP, Zhou Y, Lu SH. A novel tumor suppressor gene ECRG4 interacts directly with TMPRSS11A (ECRG1) to inhibit cancer cell growth in esophageal carcinoma. BMC Cancer. 2011;11:52.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Vanaja DK, Ehrich M, Van den Boom D, Cheville JC, Karnes RJ, Tindall DJ, et al. Hypermethylation of genes for diagnosis and risk stratification of prostate cancer. Cancer Invest. 2009;27:549–60.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Camões MJ, Paulo P, Ribeiro FR, Barros-Silva JD, Almeida M, Costa VL, et al. Potential downstream target genes of aberrant ETS transcription factors are differentially affected in Ewing’s sarcoma and prostate carcinoma. PLoS One. 2012;7(11):e49819.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Gotze S, Feldhaus V, Traska T, Wolter M, Reifenberger G, Tannapfel A, et al. ECRG4 is a candidate tumor suppressor gene frequently hypermethylated in colorectal carcinoma and glioma. BMC Cancer. 2009;9:447.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Li W, Liu XR, Zhang B, Qi DX, Zhang LH, Jin YH, et al. Overexpression of candidate tumor suppressor ECRG4 inhibits glioma proliferation and invasion. J Exp Clin Cancer Res. 2010;29:89.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Wang YB, Ba CF. Promoter methylation of esophageal cancer-related gene 4 in gastric cancer tissue and its clinical significance. Hepatogastroenterology. 2012;59(118):1696–8.PubMedGoogle Scholar
  10. 10.
    Qu Y, Dang S, Hou P. Gene methylation in gastric cancer. Clin Chim Acta. 2013;424:53–65.CrossRefPubMedGoogle Scholar
  11. 11.
    Aschebrook-Kilfoy B, Ward MH, Sabra MM, Devesa SS. Thyroid cancer incidence patterns in the United States by histologic type, 1992–2006. Thyroid. 2011;21(2):125–34.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Wang N, Dong CR, Jiang R, Tang C, Yang L, Jiang QF, et al. Overexpression of HIF-1α, metallothionein and SLUG is associated with high TNM stage and lymph node metastasis in papillary thyroid carcinoma. Int J Clin Exp Pathol. 2013;7(1):322–30.PubMedPubMedCentralGoogle Scholar
  13. 13.
    Siironen P, Nordling S, Louhimo J, Haapiainen R, Haglund C. Immunohistochemical expression of Bcl-2, Ki-67, and p21 in patients with papillary thyroid cancer. Tumour Biol. 2005;26(1):50–6.CrossRefPubMedGoogle Scholar
  14. 14.
    Hamzany Y, Soudry E, Strenov Y, Lipschitz N, Segal K, Hadar T, et al. Early death from papillary thyroid carcinoma. Am J Otolaryngol. 2012;33(1):104–8.CrossRefPubMedGoogle Scholar
  15. 15.
    Herman JG, Baylin SB. Gene silencing in cancer in association with promoter hypermethylation. N Engl J Med. 2003;349:2042–54.CrossRefPubMedGoogle Scholar
  16. 16.
    Pattani KM, Soudry E, Glazer CA, Ochs MF, Wang H, Schussel J, et al. MAGEB2 is activated by promoter demethylation in head and neck squamous cell carcinoma. PLoS One. 2012;7(9):e45534.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Kim JY, Kim KB, Eom GH, Choe N, Kee HJ, Son HJ, et al. KDM3B is the H3K9 demethylase involved in transcriptional activation of lmo2 in leukemia. Mol Cell Biol. 2012;32(14):2917–33.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Su T, Liu H, Lu S. Cloning and identification of cDNA fragments related to human esophageal cancer. Zhonghua Zhong Liu Za Zhi. 1998;20:254–7.PubMedGoogle Scholar
  19. 19.
    Yue CM, Deng DJ, Bi MX, et al. Expression of ECRG4, a novel esophageal cancer-related gene, downregulated by CpG island hypermethylation in human esophageal squamous cell carcinoma. World J Gastroenterol. 2003;9(6):1174–8.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Dang X, Podvin S, Coimbra R, Eliceiri B, Baird A. Cell-specific processing and release of the hormone-like precursor and candidate tumor suppressor gene product, Ecrg4. Cell Tissue Res. 2012;348(3):505–14.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  1. 1.Department of Laboratory Medicine, School of MedicineTaizhou UniversityTaizhouChina
  2. 2.Department of Clinical LaboratoryTaizhou Municipal HospitalTaizhouChina
  3. 3.Blood Center of WisconsinMilwaukeeUSA
  4. 4.Department of Pathology, School of MedicineZhejiang UniversityHangzhouChina

Personalised recommendations