Digestive Diseases and Sciences

, Volume 59, Issue 2, pp 328–335 | Cite as

HER2 mRNA Status Contributes to the Discrepancy Between Gene Amplification and Protein Overexpression in Gastric Cancer

  • Gui-Fen Ma
  • Yi-Mei Liu
  • Hong Gao
  • Qing Miao
  • Tian-Cheng Luo
  • Xiao-Qing Zeng
  • Shi-Yao Chen
Original Article
First Online:
Received:
Accepted:

Abstract

Background

Human epidermal growth factor receptor 2 (HER2) is an important proto-oncogene of prognostic use in gastric cancer (GC). Fluorescence in-situ hybridization (FISH) and immunohistochemistry (IHC) are the main clinical methods of detection of HER2, but consistency between the methods is poor and the cause of the discrepancy is unclear.

Aim

To investigate the involvement of HER2 mRNA status in the disparity between gene amplification and protein overexpression.

Methods

We investigated HER2 gene, mRNA, and protein profiles in gastric precancer and cancer tissues by use of the molecular approaches FISH, real-time polymerase chain reaction, and IHC. The relationships between HER2 and matrix metalloproteinase 9 (MMP9) and Smad7 expression were analyzed and the involvement of HER2 in the interaction between tumor cells and lymphocytes was investigated by coculturing GC cell lines with peripheral blood mononuclear cells (PBMCs).

Results

HER2 protein expression was significantly increased in cancer compared with precancer (P = 0.003), and the corresponding mRNA levels were significantly lower in precancer and cancer tissues than in normal tissues (κ = 0.290, P = 0.025). HER2 mRNA levels were significantly higher in tumor than in peritumor tissue (P = 0.028), and were positively correlated with MMP9 and Smad7 mRNA levels in tumor tissues. HER2 mRNA expression in GC cell lines was increased by coculture with PBMCs.

Conclusions

Different HER2 mRNA profiles, possibly in relation to contact between tumor cells and lymphocytes, might help to explain the discrepancy between gene amplification and protein overexpression results.

Keywords

HER2 Gastric cancer Matrix metalloproteinase 9 Smad7 Coculture 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgments

Research Fund of Zhongshan Hospital (2013ZSQN08).

Conflict of interest

None.

Supplementary material

10620_2013_2925_MOESM1_ESM.doc (39 kb)
Supplementary material 1 (DOC 39 kb)

References

  1. 1.
    Akiyama T, Sudo C, Ogawara H, Toyoshima K, Yamamoto T. The product of the human c-erbB-2 gene: a 185-kilodalton glycoprotein with tyrosine kinase activity. Science. 1986;232:1644–1646.PubMedCrossRefGoogle Scholar
  2. 2.
    Borg A, Linell F, Idvall I, et al. HER2/neu amplification and comedo type breast carcinoma. Lancet. 1989;1:1268–1269.PubMedCrossRefGoogle Scholar
  3. 3.
    Kesisis G, Kontovinis LF, Gennatas K, Kortsaris AH. Biological markers in breast cancer prognosis and treatment. J BUON. 2010;15:447–454.PubMedGoogle Scholar
  4. 4.
    Lorenzen S, Lordick F. How will human epidermal growth factor receptor 2-neu data impact clinical management of gastric cancer? Curr Opin Oncol. 2011;23:396–402.PubMedCrossRefGoogle Scholar
  5. 5.
    Egeblad M, Werb Z. New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer. 2002;2:161–174.PubMedCrossRefGoogle Scholar
  6. 6.
    Albarello L, Pecciarini L, Doglioni C. HER2 testing in gastric cancer. Adv Anat Pathol. 2011;18:53–59.PubMedCrossRefGoogle Scholar
  7. 7.
    Sornmayura P, Rerkamnuaychoke B, Jinawath A, Euanorasetr C. Human epidermal growth-factor receptor 2 overexpression in gastric carcinoma in Thai patients. J Med Assoc Thai. 2012;95:88–95.PubMedGoogle Scholar
  8. 8.
    Bang YJ, Van Cutsem E, Feyereislova A, et al. Trastuzumab in combination with chemotherapy versus chemotherapy alone for treatment of HER2-positive advanced gastric or gastro-oesophageal junction cancer (ToGA): a phase 3, open-label, randomised controlled trial. Lancet. 2010;376:687–697.PubMedCrossRefGoogle Scholar
  9. 9.
    Arnold S, Mira E, Muneer S, et al. Forced expression of MMP9 rescues the loss of angiogenesis and abrogates metastasis of pancreatic tumors triggered by the absence of host sparc. Exp Biol Med (Maywood). 2008;233:860–873.CrossRefGoogle Scholar
  10. 10.
    Kim YH, Lee HS, Lee HJ, et al. Prognostic significance of the expression of Smad4 and Smad7 in human gastric carcinomas. Ann Oncol. 2004;15:574–580.PubMedCrossRefGoogle Scholar
  11. 11.
    Heikkinen PT, Nummela M, Jokilehto T, Grenman R, Kahari VM, Jaakkola PM. Hypoxic conversion of SMAD7 function from an inhibitor into a promoter of cell invasion. Cancer Res. 2010;70:5984–5993.PubMedCrossRefGoogle Scholar
  12. 12.
    Kim S, Han J, Lee SK, et al. Smad7 acts as a negative regulator of the epidermal growth factor (EGF) signaling pathway in breast cancer cells. Cancer Lett. 2012;314:147–154.PubMedCrossRefGoogle Scholar
  13. 13.
    Yannelli JR, Tucker JA, Hidalgo G, Perkins S, Kryscio R, Hirschowitz EA. Characteristics of PBMC obtained from leukapheresis products and tumor biopsies of patients with non-small cell lung cancer. Oncol Rep. 2009;22:1459–1471.PubMedCrossRefGoogle Scholar
  14. 14.
    Nowak M, Klink M, Glowacka E, et al. Production of cytokines during interaction of peripheral blood mononuclear cells with autologous ovarian cancer cells or benign ovarian tumour cells. Scand J Immunol. 2010;71:91–98.PubMedCrossRefGoogle Scholar
  15. 15.
    Poggi A, Zocchi MR. Mechanisms of tumor escape: role of tumor microenvironment in inducing apoptosis of cytolytic effector cells. Arch Immunol Ther Exp (Warsz). 2006;54:323–333.CrossRefGoogle Scholar
  16. 16.
    Minot DM, Voss J, Rademacher S, et al. Image analysis of HER2 immunohistochemical staining. Reproducibility and concordance with fluorescence in situ hybridization of a laboratory-validated scoring technique. Am J Clin Pathol. 2012;137:270–276.PubMedCrossRefGoogle Scholar
  17. 17.
    Ruschoff J, Dietel M, Baretton G, et al. HER2 diagnostics in gastric cancer-guideline validation and development of standardized immunohistochemical testing. Virchows Arch. 2010;457:299–307.PubMedCentralPubMedCrossRefGoogle Scholar
  18. 18.
    Ma GF, Miao Q, Zeng XQ, et al. Transforming growth factor-b1 and -b2 in gastric precancer and cancer and roles in tumor-cell interactions with peripheral blood mononuclear cells in vitro. PLoS One. 2013;8:e54249.PubMedCentralPubMedCrossRefGoogle Scholar
  19. 19.
    Cuadros M, Villegas R. Systematic review of HER2 breast cancer testing. Appl Immunohistochem Mol Morphol. 2009;17:1–7.PubMedCrossRefGoogle Scholar
  20. 20.
    Hofmann M, Stoss O, Shi D, et al. Assessment of a HER2 scoring system for gastric cancer: results from a validation study. Histopathology. 2008;52:797–805.PubMedCrossRefGoogle Scholar
  21. 21.
    Sauter G, Lee J, Bartlett JM, Slamon DJ, Press MF. Guidelines for human epidermal growth factor receptor 2 testing: biologic and methodologic considerations. J Clin Oncol. 2009;27:1323–1333.PubMedCrossRefGoogle Scholar
  22. 22.
    Vanderhaegen J, Paridaens R, Neven P. Adjuvant trastuzumab in HER2-positive breast cancer. N Engl J Med. 2012;366:664–666.PubMedGoogle Scholar
  23. 23.
    Muller BM, Kronenwett R, Hennig G, et al. Quantitative determination of estrogen receptor, progesterone receptor, and HER2 mRNA in formalin-fixed paraffin-embedded tissue—a new option for predictive biomarker assessment in breast cancer. Diagn Mol Pathol. 2011;20:1–10.PubMedCrossRefGoogle Scholar
  24. 24.
    Iverson AA, Gillett C, Cane P, et al. A single-tube quantitative assay for mRNA levels of hormonal and growth factor receptors in breast cancer specimens. J Mol Diagn. 2009;11:117–130.PubMedCentralPubMedCrossRefGoogle Scholar
  25. 25.
    Moerland E, van Hezik RL, van der Aa TC, van Beek MW, van den Brule AJ. Detection of HER2 amplification in breast carcinomas: comparison of multiplex ligation-dependent probe amplification (MLPA) and fluorescence in situ hybridization (FISH) combined with automated spot counting. Cell Oncol. 2006;28:151–159.PubMedGoogle Scholar
  26. 26.
    Tubbs RR, Pettay JD, Roche PC, Stoler MH, Jenkins RB, Grogan TM. Discrepancies in clinical laboratory testing of eligibility for trastuzumab therapy: apparent immunohistochemical false-positives do not get the message. J Clin Oncol. 2001;19:2714–2721.PubMedGoogle Scholar
  27. 27.
    Kuesters S, Maurer M, Burger AM, Metz T, Fiebig HH. Correlation of ErbB2 gene status, mRNA and protein expression in a panel of >100 human tumor xenografts of different origin. Onkologie. 2006;29:249–256.PubMedCrossRefGoogle Scholar
  28. 28.
    Tanner M, Hollmen M, Junttila TT, et al. Amplification of HER-2 in gastric carcinoma: association with Topoisomerase IIalpha gene amplification, intestinal type, poor prognosis and sensitivity to trastuzumab. Ann Oncol. 2005;16:273–278.PubMedCrossRefGoogle Scholar
  29. 29.
    Moelans CB, van Diest PJ, Milne AN, Offerhaus GJ. Her-2/neu testing and therapy in gastroesophageal adenocarcinoma. Patholog Res Int. 2011;2011:674182.PubMedCentralGoogle Scholar
  30. 30.
    Pellikainen JM, Ropponen KM, Kataja VV, Kellokoski JK, Eskelinen MJ, Kosma VM. Expression of matrix metalloproteinase (MMP)-2 and MMP-9 in breast cancer with a special reference to activator protein-2, HER2, and prognosis. Clin Cancer Res. 2004;10:7621–7628.PubMedCrossRefGoogle Scholar
  31. 31.
    O-Charoenrat P, Rhys-Evans P, Eccles S. A synthetic matrix metalloproteinase inhibitor prevents squamous carcinoma cell proliferation by interfering with epidermal growth factor receptor autocrine loops. Int J Cancer. 2002;100:527–533.PubMedCrossRefGoogle Scholar
  32. 32.
    Asrani K, Keri RA, Galisteo R, et al. The HER2- and heregulin beta1 (Hrg)-inducible TNFR superfamily member Fn14 promotes HRG-driven breast cancer cell migration, invasion, and MMP9 expression. Mol Cancer Res. 2013;11:393–404.PubMedCrossRefGoogle Scholar
  33. 33.
    Jang JY, Jeon YK, Kim CW. Degradation of HER2/neu by ANT2 shRNA suppresses migration and invasiveness of breast cancer cells. BMC Cancer. 2010;10:391.PubMedCentralPubMedCrossRefGoogle Scholar
  34. 34.
    Leng A, Liu T, He Y, Li Q, Zhang G. Smad4/Smad7 balance: a role of tumorigenesis in gastric cancer. Exp Mol Pathol. 2009;87:48–53.Google Scholar
  35. 35.
    Dowdy SC, Mariani A, Janknecht R. HER2/Neu- and TAK1-mediated up-regulation of the transforming growth factor beta inhibitor Smad7 via the ETS protein ER81. J Biol Chem. 2003;278:44377–44384.PubMedCrossRefGoogle Scholar
  36. 36.
    Lu X, Liu J, Li H, et al. Conversion of intratumoral regulatory t cells by human gastric cancer cells is dependent on transforming growth factor-beta1. J Surg Oncol. 2011;104:571–577.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Gui-Fen Ma
    • 1
  • Yi-Mei Liu
    • 1
  • Hong Gao
    • 1
  • Qing Miao
    • 1
  • Tian-Cheng Luo
    • 1
  • Xiao-Qing Zeng
    • 1
  • Shi-Yao Chen
    • 1
    • 2
  1. 1.Department of Gastroenterology, Zhongshan HospitalFudan UniversityShanghaiChina
  2. 2.Endoscopy Center, Zhongshan HospitalFudan UniversityShanghaiChina

Personalised recommendations