Molecular and Cellular Biochemistry

, Volume 366, Issue 1–2, pp 223–229 | Cite as

Overexpression of ZEB1 associated with metastasis and invasion in patients with gastric carcinoma



The aim of this study was to investigate the expression of ZEB1 in gastric carcinoma, its correlation with the clinicopathology of gastric carcinoma, and the role of ZEB1 in invasion and metastasis in gastric carcinoma. ZEB1 expression was analyzed by immunohistochemistry and Western blot in 45 gastric carcinoma tissue samples that contained the adjacent gastric mucosa. The correlation between ZEB1 expression, the occurrence and development of gastric cancer, and clinical pathology was investigated. ZEB1 expression in the human gastric carcinoma cell line AGS was downregulated by RNA interference, and changes in ZEB1 expression corresponded with changes in the invasive and metastatic ability of AGS cells. Immunohistochemistry revealed that ZEB1 protein expression in gastric carcinoma tissues was significantly higher than in normal gastric mucosa tissues (p < 0.001). A lower degree of differentiation of gastric cancer (p = 0.009), a higher TNM (tumor, node, and metastasis) stage (p = 0.010), and a larger scope of invasion were correlated with higher expression of ZEB1 (p = 0.041, 0.002). However, the expression of ZEB1 in gastric carcinoma tissue was independent of gender, age, and tumor size (p > 0.05). Western blot results also showed that ZEB1 protein expression was significantly higher in gastric carcinoma tissue than in the adjacent normal gastric mucosa tissue (p = 0.008). A lower degree of differentiation of the gastric carcinoma correlated with a higher TNM stage, and a larger scope of invasion correlated with increased ZEB1 expression (p = 0.023). Transfection of ZEB1 siRNA in AGS cells significantly decreased the expression level of ZEB1 protein (p = 0.035). Furthermore, the number of cells that could pass through the Transwell chamber was significantly lower in the transfected group than in the non-transfected control group (p = 0.039), indicating that the suppression of ZEB1 expression could significantly reduce the invasive and metastatic ability of AGS cells (p = 0.005). Concluding, in gastric carcinoma tissue, overexpression of ZEB1 may be related to the occurrence and development as well as invasion and metastasis of gastric carcinoma.


Gastric carcinoma Metastasis ZEB1 RNAi 


Conflict of interest



  1. 1.
    Yang D, Hendifar A, Lenz C, Togawa K, Lenz F, Lurje G, Pohl A, Winder T, Ning Y, Groshen S, Lenz H (2011) Survival of metastatic gastric cancer: significance of age, sex and race/ethnicity. J Gastrointest Oncol 2:77–84Google Scholar
  2. 2.
    Jimenez P, Pathak A, Phan A (2011) The role of taxanes in the management of gastroesphageal cancer. J Gastrointest Oncol 2:240–249Google Scholar
  3. 3.
    Tan K, Quek T, Wong N, Li K, Lim K (2011) Emergency surgery for perforated gastric malignancy: an institution’s experience and review of the literature. J Gastrointest Oncol 2:13–18Google Scholar
  4. 4.
    Saglam S, Aykan N, Sakar B, Gulluoglu M, Balik E, Karanlik H (2011) A pilot study evaluating the safety and toxicity of epirubicin, cisplatin, and UFT (ECU regimen) in advanced gastric carcinoma. J Gastrointest Oncol 2:19–26Google Scholar
  5. 5.
    Hopkins S, Yang G (2011) FDG PET imaging in the staging and management of gastric cancer. J Gastrointest Oncol 2:39–44Google Scholar
  6. 6.
    Remacle JE, Kraft H, Lerchner W, Wuytens G, Collart C, Verschueren K, Smith JC, Huylebroeck D (1999) New mode of DNA binding of multi-zinc finger transcription factors: deltaEF1 family members bind with two hands to two target sites. EMBO J 18:5073–5084PubMedCrossRefGoogle Scholar
  7. 7.
    Vandewalle C, Comijn J, De Craene B, Vermassen P, Bruyneel E, Andersen H, Tulchinsky E, Van Roy F, Berx G (2005) SIP1/ZEB2 induces EMT by repressing genes of different epithelial cell–cell junctions. Nucleic Acids Res 33:6566–6578PubMedCrossRefGoogle Scholar
  8. 8.
    Postigo AA, Depp JL, Taylor JJ, Kroll KL (2003) Regulation of Smad signaling through a differential recruitment of coactivators and corepressors by ZEB proteins. EMBO J 22:2453–2462PubMedCrossRefGoogle Scholar
  9. 9.
    Fontemaggi G, Gurtner A, Strano S, Higashi Y, Sacchi A, Piaggio G, Blandino G (2001) The transcriptional repressor ZEB regulates p73 expression at the crossroad between proliferation and differentiation. Mol Cell Biol 21:8461–8470PubMedCrossRefGoogle Scholar
  10. 10.
    Stridh P, Thessen Hedreul M, Beyeen AD, Adzemovic MZ, Laaksonen H, Gillett A, Ockinger J, Marta M, Lassmann H, Becanovic K, Jagodic M, Olsson T (2010) Fine-mapping resolves Eae23 into two QTLs and implicates ZEB1 as a candidate gene regulating experimental neuroinflammation in rat. PLoS ONE 5:e12716PubMedCrossRefGoogle Scholar
  11. 11.
    Hidaka T, Nakahata S, Hatakeyama K, Hamasaki M, Yamashita K, Kohno T, Arai Y, Taki T, Nishida K, Okayama A, Asada Y, Yamaguchi R, Tsubouchi H, Yokota J, Taniwaki M, Higashi Y, Morishita K (2008) Down-regulation of TCF8 is involved in the leukemogenesis of adult T-cell leukemia/lymphoma. Blood 112:383–393PubMedCrossRefGoogle Scholar
  12. 12.
    Postigo AA, Dean DC (1999) Independent repressor domains in ZEB regulate muscle and T-cell differentiation. Mol Cell Biol 19:7961–7971PubMedGoogle Scholar
  13. 13.
    Williams TM, Moolten D, Burlein J, Romano J, Bhaerman R, Godillot A, Mellon M, Rauscher FJ 3rd, Kant JA (1991) Identification of a zinc finger protein that inhibits IL-2 gene expression. Science 254:1791–1794PubMedCrossRefGoogle Scholar
  14. 14.
    Putzke AP, Ventura AP, Bailey AM, Akture C, Opoku-Ansah J, Celiktaş M, Hwang MS, Darling DS, Coleman IM, Nelson PS, Nguyen HM, Corey E, Tewari M, Morrissey C, Vessella RL, Knudsen BS (2011) Metastatic progression of prostate cancer and e-cadherin regulation by zeb1 and SRC family kinases. Am J Pathol 179:400–410PubMedCrossRefGoogle Scholar
  15. 15.
    Jensen K, Afroze S, Munshi MK, Guerrier M, Glaser SS (2012) Mechanisms for nicotine in the development and progression of gastrointestinal cancers. Transl Gastrointest Cancer. doi: 10.3978/j.issn.2224-4778.2011.12.01
  16. 16.
    Lorenzatti G, Huang W, Pal A, Cabanillas AM, Kleer CG (2011) CCN6 (WISP3) decreases ZEB1-mediated EMT and invasion by attenuation of IGF-1 receptor signaling in breast cancer. J Cell Sci 124:1752–1758PubMedCrossRefGoogle Scholar
  17. 17.
    Bao B, Wang Z, Ali S, Kong D, Li Y, Ahmad A, Banerjee S, Azmi AS, Miele L, Sarkar FH (2011) Notch-1 induces epithelial-mesenchymal transition consistent with cancer stem cell phenotype in pancreatic cancer cells. Cancer Lett 307:26–36PubMedCrossRefGoogle Scholar
  18. 18.
    Roy BC, Kohno T, Iwakawa R, Moriguchi T, Kiyono T, Morishita K, Sanchez-Cespedes M, Akiyama T, Yokota J (2010) Involvement of LKB1 in epithelial-mesenchymal transition (EMT) of human lung cancer cells. Lung Cancer 70:136–145PubMedCrossRefGoogle Scholar
  19. 19.
    Leshem O, Madar S, Kogan-Sakin I, Kamer I, Goldstein I, Brosh R, Cohen Y, Jacob-Hirsch J, Ehrlich M, Ben-Sasson S, Goldfinger N, Loewenthal R, Gazit E, Rotter V, Berger R (2011) TMPRSS2/ERG promotes epithelial to mesenchymal transition through the ZEB1/ZEB2 axis in a prostate cancer model. PLoS ONE 6:e21650PubMedCrossRefGoogle Scholar
  20. 20.
    Zhu XL, Liang L, Ding YQ (2008) Overexpression of FMNL2 is closely related to metastasis of colorectal cancer. Int J Colorectal Dis 23:1041–1047PubMedCrossRefGoogle Scholar
  21. 21.
    Vora HH, Patel NA, Rajvik KN, Mehta SV, Brahmbhatt BV, Shah MJ, Shukla SN, Shah PM (2009) Cytokeratin and vimentin expression in breast cancer. Int J Biol Markers 24:38–46PubMedGoogle Scholar
  22. 22.
    Murray D, Precht P, Balakir R, Horton WE Jr (2000) The transcription factor deltaEF1 is inversely expressed with type II collagen mRNA and can repress Col2a1 promoter activity in transfected chondrocytes. J Biol Chem 275:3610–3618PubMedCrossRefGoogle Scholar
  23. 23.
    Dohadwala M, Yang SC, Luo J, Sharma S, Batra RK, Huang M, Lin Y, Goodglick L, Krysan K, Fishbein MC, Hong L, Lai C, Cameron RB, Gemmill RM, Drabkin HA, Dubinett SM (2006) Cyclooxygenase-2-dependent regulation of E-cadherin: prostaglandin E(2) induces transcriptional repressors ZEB1 and snail in non-small cell lung cancer. Cancer Res 66:5338–5345PubMedCrossRefGoogle Scholar
  24. 24.
    Manavella PA, Roqueiro G, Darling DS, Cabanillas AM (2007) The ZFHX1A gene is differentially autoregulated by its isoforms. Biochem Biophys Res Commun 360:621–626PubMedCrossRefGoogle Scholar
  25. 25.
    Postigo AA (2003) Opposing functions of ZEB proteins in the regulation of the TGFbeta/BMP signaling pathway. EMBO J 22:2443–2452PubMedCrossRefGoogle Scholar
  26. 26.
    Nakahata S, Yamazaki S, Nakauchi H, Morishita K (2010) Downregulation of ZEB1 and overexpression of Smad7 contribute to resistance to TGF-beta1-mediated growth suppression in adult T-cell leukemia/lymphoma. Oncogene 29:4157–4169PubMedCrossRefGoogle Scholar
  27. 27.
    Bateman G, Ricketts DN, Saunders WP (2003) Fibre-based post systems: a review. Br Dent J 195:43–48PubMedCrossRefGoogle Scholar
  28. 28.
    Yu HG, Li JY, Yang YN, Luo HS, Yu JP, Meier JJ, Schrader H, Bastian A, Schmidt WE, Schmitz F (2003) Increased abundance of cyclooxygenase-2 correlates with vascular endothelial growth factor-A abundance and tumor angiogenesis in gastric cancer. Cancer Lett 195:43–51PubMedCrossRefGoogle Scholar
  29. 29.
    Qian Y, Corum L, Meng Q, Blenis J, Zheng JZ, Shi X, Flynn DC, Jiang BH (2004) PI3 K induced actin filament remodeling through Akt and p70S6K1: implication of essential role in cell migration. Am J Physiol Cell Physiol 286:C153–C163PubMedCrossRefGoogle Scholar
  30. 30.
    Livraghi T (2001) Role of percutaneous ethanol injection in the treatment of hepatocellular carcinoma. Dig Dis 19:292–300PubMedCrossRefGoogle Scholar
  31. 31.
    Eger A, Aigner K, Sonderegger S, Dampier B, Oehler S, Schreiber M, Berx G, Cano A, Beug H, Foisner R (2005) DeltaEF1 is a transcriptional repressor of E-cadherin and regulates epithelial plasticity in breast cancer cells. Oncogene 24:2375–2385PubMedCrossRefGoogle Scholar
  32. 32.
    van Roy F, Berx G (2008) The cell–cell adhesion molecule E-cadherin. Cell Mol Life Sci 65:3756–3788PubMedCrossRefGoogle Scholar
  33. 33.
    Rachagani S, Senapati S, Chakraborty S, Ponnusamy MP, Kumar S, Smith LM, Jain M, Batra SK (2011) Activated Kras(G12D) is associated with invasion and metastasis of pancreatic cancer cells through inhibition of E-cadherin. Br J Cancer 104:1038–1048PubMedCrossRefGoogle Scholar
  34. 34.
    Erdem H, Gundogdu C, Sipal S (2011) Correlation of E-cadherin, VEGF, COX-2 expression to prognostic parameters in papillary thyroid carcinoma. Exp Mol Pathol 90:312–317PubMedCrossRefGoogle Scholar
  35. 35.
    Saad AA, Awed NM, Abd Elkerim NN, El-Shennawy D, Alfons MA, Elserafy ME, Darwish YW, Barakat EM, Ezz-Elarab SS (2010) Prognostic significance of E-cadherin expression and peripheral blood micrometastasis in gastric carcinoma patients. Ann Surg Oncol 17:3059–3067PubMedCrossRefGoogle Scholar
  36. 36.
    Bae KM, Parker NN, Dai Y, Vieweg J, Siemann DW (2011) E-cadherin plasticity in prostate cancer stem cell invasion. Am J Cancer Res 1:71–84PubMedGoogle Scholar
  37. 37.
    Rajeshwari KV, Lata K, Pant DC, Kishore VV (2001) A novel process using enhanced acidification and a UASB reactor for biomethanation of vegetable market waste. Waste Manag Res 19:292–300PubMedCrossRefGoogle Scholar
  38. 38.
    Potiron VA, Sharma G, Nasarre P, Clarhaut JA, Augustin HG, Gemmill RM, Roche J, Drabkin HA (2007) Semaphorin SEMA3F affects multiple signaling pathways in lung cancer cells. Cancer Res 67:8708–8715PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2012

Authors and Affiliations

  • Baoqing Jia
    • 1
  • Hongyi Liu
    • 1
  • Qinglong Kong
    • 1
  • Bing Li
    • 1
  1. 1.Department of Surgical OncologyGeneral Hospital of Chinese People’s Liberation ArmyBeijingPeople’s Republic of China

Personalised recommendations