Digestive Diseases and Sciences

, Volume 58, Issue 3, pp 715–723 | Cite as

DNA Methylation of NDRG2 in Gastric Cancer and Its Clinical Significance

  • Xiaojing Chang
  • Zhenhua Li
  • Jinguo Ma
  • Peng Deng
  • Shuanglong Zhang
  • Yu Zhi
  • Jing Chen
  • Dongqiu Dai
Original Article



Gastric cancer is one of the most common digestive malignancies worldwide. N-myc downstream-regulated gene 2 (NDRG2) is a differentiation-related gene that is considered to be a metastasis suppressor gene. In this study, we examined the expression and DNA methylation of NDRG2 in gastric cancer cell lines and tissues, as well as its clinical significance.


Six gastric cancer cell lines and 42 paired normal and gastric cancer tissue samples were used to assess NDRG2 mRNA expression using RT-PCR. NDRG2 DNA methylation status was evaluated by methylation-specific PCR (MSP) in gastric cancer cell lines and tissues. The suppression of NDRG2 in BGC823 cells by siRNA transfection was utilized to detect the role of NDRG2 in gastric cancer progression.


NDRG2 mRNA was down-regulated in gastric cancer cell lines and tissues, and its expression was just related to lymph node metastasis (p = 0.032). MSP showed methylation of NDRG2 in 54.0 % (47/87) of primary gastric cancer specimens and in 20.0 % (16/80) of corresponding nonmalignant gastric tissues. NDRG2 methylation was related to depth of tumor invasion, Borrmann classification and TNM stage (p < 0.05). Upon treatment with 5-aza-2′-deoxycytidine and trichostatin A, NDRG2 expression was upregulated in HGC27 cells, and demethylation of the highly metastatic cell line, MKN45, inhibited cell invasion. Furthermore, the suppression of NDRG2 by siRNA transfection enhanced BGC823 cells invasion.


Our results suggest that the aberrant methylation of NDRG2 may be mainly responsible for its downregulation in gastric cancer, and may play an important role in the metastasis of gastric cancer.


Differentiation Gastric carcinoma DNA methylation Histone modification 


  1. 1.
    Leung WK, Wu MS, Kakugawa Y, et al. Screening for gastric cancer in Asia: current evidence and practice. Lancet Oncol. 2008;9:279–287.PubMedCrossRefGoogle Scholar
  2. 2.
    Melotte V, Qu X, Ongenaert M, et al. The N-myc downstream regulated gene (NDRG) family: diverse functions, multiple applications. FASEB J. 2010;24:4153–4166.PubMedCrossRefGoogle Scholar
  3. 3.
    Shi H, Jin H, Chu D, et al. Suppression of N-myc downstream-regulated gene 2 is associated with induction of Myc in colorectal cancer and correlates closely with differentiation. Biol Pharm Bull. 2009;32:968–975.PubMedCrossRefGoogle Scholar
  4. 4.
    Hu XL, Liu XP, Deng YC, et al. Expression analysis of the NDRG2 gene in mouse embryonic and adult tissues. Cell Tiss Res. 2006;325:67–76.CrossRefGoogle Scholar
  5. 5.
    Sun Z, Shen L, Sun X, et al. Variation of NDRG2 and c-Myc expression in rat heart during the acute stage of ischemia/reperfusion injury. Histochem Cell Biol. 2011;135:27–35.PubMedCrossRefGoogle Scholar
  6. 6.
    Mordalska A, Latek J, Ferenc T, et al. Evaluation of NDRG2 gene expression in primary papillary thyroid carcinoma and in metastases of this neoplasm to regional lymph nodes. Thyroid Res. 2010;3:6.PubMedCrossRefGoogle Scholar
  7. 7.
    Ma JJ, Liao CG, Jiang X, Zhao HD, Yao LB, Bao TY. NDRG2 suppresses the proliferation of clear cell renal cell carcinoma cell A-498. J Exp Clin Cancer Res. 2010;29:103.PubMedCrossRefGoogle Scholar
  8. 8.
    Gao L, Wu GJ, Liu XW, et al. Suppression of invasion and metastasis of prostate cancer cells by overexpression of NDRG2 gene. Cancer Lett. 2011;310:94–100.PubMedCrossRefGoogle Scholar
  9. 9.
    Choi SC, Yoon SR, Park YP, et al. Expression of NDRG2 is related to tumor progression and survival of gastric cancer patients through Fas-mediated cell death. Exp Mol Med. 2007;39:705–714.PubMedCrossRefGoogle Scholar
  10. 10.
    Zheng J, Liu Q, Li Y, et al. NDRG2 expression regulates CD24 and metastatic potential of breast cancer cells. Asian Pac J Cancer Prev. 2010;11:1817–1821.PubMedGoogle Scholar
  11. 11.
    Zheng J, Li Y, Yang J, et al. NDRG2 inhibits hepatocellular carcinoma adhesion, migration and invasion by regulating CD24 expression. BMC Cancer. 2011;11:1–9.PubMedCrossRefGoogle Scholar
  12. 12.
    Zhang J, Liu J, Li X, et al. The physical and functional interaction of NDRG2 with MSP58 in cells. Biochem Biophys Res Commun. 2007;352:6–11.PubMedCrossRefGoogle Scholar
  13. 13.
    Furuta H, Kondo Y, Nakahata S, Hamasaki M, Sakoda S, Morishita K. NDRG2 is a candidate tumor-suppressor for oral squamous-cell carcinoma. Biochem Biophys Res Commun. 2010;391:1785–1791.PubMedCrossRefGoogle Scholar
  14. 14.
    Tepel M, Roerig P, Wolter M, et al. Frequent promoter hypermethylation and transcriptional downregulation of the NDRG2 gene at 14q11.2 in primary glioblastoma. Int J Cancer. 2008;123:2080–2086.PubMedCrossRefGoogle Scholar
  15. 15.
    Liu N, Wang L, Liu X, et al. Promoter methylation, mutation, and genomic deletion are involved in the decreased NDRG2 expression levels in several cancer cell lines. Biochem Biophys Res Commun. 2007;358:164–169.PubMedCrossRefGoogle Scholar
  16. 16.
    Piepoli A, Cotugno R, Merla G, et al. Promoter methylation correlates with reduced NDRG2 expression in advanced colon tumor. BMC Med Genomics. 2009;2:11.PubMedCrossRefGoogle Scholar
  17. 17.
    Assämäki R, Sarlomo-Rikala M, Lopez-Guerrero JA, et al. Array comparative genomic hybridization analysis of chromosomal imbalances and their target genes in gastrointestinal stromal tumors. Genes Chromosom Cancer. 2007;46:564–576.PubMedCrossRefGoogle Scholar
  18. 18.
    Ma J, Liu WC, Yan XH, et al. Inhibition of endothelial cell proliferation and tumor angiogenesis by up-regulating NDRG2 expression in breast cancer cells. PLoS One. 2012;7:e32368.PubMedCrossRefGoogle Scholar
  19. 19.
    Shen WJ, Dai DQ, Teng Y, Liu HB. Regulation of demethylation and re-expression of RASSF1A gene in gastric cancer cell lines by combined treatment of 5-Aza-CdR and NaB. World J Gastroenterol. 2008;14:595–600.PubMedCrossRefGoogle Scholar
  20. 20.
    Du YY, Dai DQ, Yang Z. Role of RECK methylation in gastric cancer and its clinical significance. World J Gastroenterol. 2010;16:904–908.PubMedCrossRefGoogle Scholar
  21. 21.
    Rizvi MM, Alam MS, Ali A, Mehdi SJ, Batra S, Mandal AK. Aberrant promoter methylation and inactivation of PTEN gene in cervical carcinoma from Indian population. J Cancer Res Clin Oncol. 2011;137:1255–1262.PubMedCrossRefGoogle Scholar
  22. 22.
    Chimonidou M, Strati A, Tzitzira A, et al. DNA methylation of tumor suppressor and metastasis suppressor genes in circulating tumor cells. Clin Chem. 2011;57:1169–1177.PubMedCrossRefGoogle Scholar
  23. 23.
    Yamashita S, Tsujino Y, Moriguchi K, Tatematsu M, Ushijima T. Chemical genomic screening for methylation-silenced genes in gastric cancer cell lines using 5-aza-2′-deoxycytidine treatment and oligonucleotide microarray. Cancer Sci. 2006;97:64–71.PubMedCrossRefGoogle Scholar
  24. 24.
    Kang GH, Lee S, Kim JS, Jung HY. Profile of aberrant CpG island methylation along multistep gastric carcinogenesis. Lab Invest. 2003;83:519–526.PubMedGoogle Scholar
  25. 25.
    Tost J. DNA methylation: an introduction to the biology and the disease-associated changes of a promising biomarker. Mol Biotechnol. 2010;44:71–81.PubMedCrossRefGoogle Scholar
  26. 26.
    Lorentzen A, Vogel LK, Lewinsky RH, et al. Expression of NDRG2 is down-regulated in high-risk adenomas and colorectal carcinoma. BMC Cancer. 2007;7:192.PubMedCrossRefGoogle Scholar
  27. 27.
    Lee DC, Kang YK, Kim WH, et al. Functional and clinical evidence for NDRG2 as a candidate suppressor of liver cancer metastasis. Cancer Res. 2008;68:4210–4220.PubMedCrossRefGoogle Scholar
  28. 28.
    Li L, Wang J, Shen X, et al. Expression and prognostic value of NDRG2 in human astrocytomas. J Neurol Sci. 2011;308:77–82.PubMedCrossRefGoogle Scholar
  29. 29.
    Liu N, Wang L, Li X, et al. N-myc downstream-regulated gene 2 is involved in p53-mediated apoptosis. Nucleic Acids Res. 2008;36:5335–5349.PubMedCrossRefGoogle Scholar
  30. 30.
    Hu XL, Liu XP, Lin SX, et al. NDRG2 expression and mutation in human liver and pancreatic cancers. World J Gastroenterol. 2004;10:3518–3521.PubMedGoogle Scholar
  31. 31.
    Das PM, Thor AD, Edgerton SM, Barry SK, Chen DF, Jones FE. Reactivation of epigenetically silenced HER4/ERBB4 results in apoptosis of breast tumor cells. Oncogene. 2010;29:5214–5219.PubMedCrossRefGoogle Scholar
  32. 32.
    Singh KP, Treas J, Tyaqi T, Gao W. DNA demethylation by 5-aza-2-deoxycytidine treatment abrogates 17 beta-estradiol-induced cell growth and restores expression of DNA repair genes in human breast cancer cells. Cancer Lett. 2012;316:62–69.PubMedCrossRefGoogle Scholar
  33. 33.
    Chiam K, Centenera MM, Butler LM, Tilley WD, Bianco-Miotto T. GSTP1 DNA methylation and expression status is indicative of 5-aza-2′-deoxycytidine efficacy in human prostate cancer cells. PLoS One. 2011;6:e25634.PubMedCrossRefGoogle Scholar
  34. 34.
    Koutsimpelas D, Pongsapich W, Heinrich U, Mann S, Mann WT, Brieger J. Promoter methylation of MGMT, MLH1 and RASSF1A tumor suppressor genes in head and neck squamous cell carcinoma: pharmacological genome demethylation reduces proliferation of head and neck squamous carcinoma cells. Oncol Rep. 2012;27:1135–1141.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Xiaojing Chang
    • 1
    • 2
  • Zhenhua Li
    • 2
  • Jinguo Ma
    • 2
  • Peng Deng
    • 2
  • Shuanglong Zhang
    • 2
  • Yu Zhi
    • 2
  • Jing Chen
    • 1
  • Dongqiu Dai
    • 1
    • 2
  1. 1.Department of Gastrointestinal Surgery, The Fourth Affiliated HospitalChina Medical UniversityShenyangChina
  2. 2.Cancer Center, The Fourth Affiliated HospitalChina Medical UniversityShenyangChina

Personalised recommendations