A single nucleotide polymorphism in the MMP-9 promoter affects tumor progression and invasive phenotype of gastric cancer

  • Shunji Matsumura
  • Naohide Oue
  • Hirofumi Nakayama
  • Yasuhiko Kitadai
  • Kazuhiro Yoshida
  • Yoshiyuki Yamaguchi
  • Kazue Imai
  • Kei Nakachi
  • Keisuke Matsusaki
  • Kazuaki Chayama
  • Wataru YasuiEmail author
Original Paper



Matrix metalloproteinase-9 (MMP-9, gelatinase B) plays a key role in cancer invasion and metastasis by degradating the extracellular matrix (ECM) and basement membrane barriers. A cytosine (C)-thymidine (T) single nucleotide polymorphism (SNP) at position –1562 in the MMP-9 promoter is reported to affect expression of this gene. The purpose of this study was to investigate the relation between the –1562 C/T polymorphism and the development and progression of gastric cancer.


The study population included 177 gastric cancer patients and 224 healthy control subjects. The SNP in the MMP-9 promoter was analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing. Genotype frequencies were compared between patients and controls, and the association of genotypes with clinicopathological features was studied.


Genotype frequencies in gastric cancer patients were similar to those in control subjects (P = 0.223). However, significant association was found between degree of tumor invasion, clinical stage, and lymphatic invasion and the MMP-9 polymorphism in gastric cancer patients (P<0.05, for each).


Our results indicate that the T allele in the MMP-9 promoter is associated with the invasive phenotype of gastric cancer.


MMP-9 SNP Gastric cancer Depth of tumor invasion Lymphatic invasion 



Matrix metalloproteinase-9


Single nucleotide polymorphism






Restriction fragment length polymorphism


Extracellular matrix


Polymerase chain reaction


Odds ratio


Confidence interval



We thank Masayoshi Takatani and Mutsumi Ueda, Department of Molecular Pathology, Hiroshima University Graduate School of Biomedical Sciences, for their excellent technical assistance and advice


  1. Baker EA, Leaper DJ (2003) The plasminogen activator and matrix metalloproteinase systems in colorectal cancer: relationship to tumour pathology. Eur J Cancer 39:981–988CrossRefPubMedGoogle Scholar
  2. Behrens P, Rothe M, Wellmann A, Krischler J, Wernert N (2001) The Ets-1 transcription factor is up-regulated together with MMP-1 and MMP-9 in the stroma of pre-invasive breast cancer. J Pathol 194:43–50CrossRefPubMedGoogle Scholar
  3. Chicoine E, Esteve PO, Robledo O, Van Themsche C, Potworowski EF, St-Pierre Y (2002) Evidence for the role of promoter methylation in the regulation of MMP-9 gene expression. Biochem Biophys Res Commun 297:765–772CrossRefPubMedGoogle Scholar
  4. Ferrand PE, Parry S, Sammel M, Macones GA, Kuivaniemi H, Romero R, Strauss JF 3rd (2002) A polymorphism in the matrix metalloproteinase-9 promoter is associated with increased risk of preterm premature rupture of membranes in African Americans. Mol Hum Reprod 8:494–501CrossRefPubMedGoogle Scholar
  5. Forget MA, Desrosiers RR, Beliveau R (1999) Physiological roles of matrix metalloproteinases; implications for tumor growth and metastasis. Can J Physiol Pharmacol 77:465–480CrossRefPubMedGoogle Scholar
  6. Gum R, Lengyel E, Juarez J, Chen JH, Sato H, Seiki M, Boyd D (1996) Stimulation of 92-kDa gelatinase B promoter activity by ras is mitogen-activated protein kinase kinase 1-independent and requires multiple transcription factor binding sites including closely spaced PEA3/ets and AP-1 sequences. J Biol Chem 271:10672–10680CrossRefPubMedGoogle Scholar
  7. Hujanen ES, Vaisanen A, Zheng A, Tryggvason K, Turpeenniemi-Hujanen T (1994) Modulation of M(r) 72,000 and M(r) 92,000 type-IV collagenase (gelatinase A and B) gene expression by interferons alpha and gamma in human melanoma. Int J Cancer 58:582–586PubMedGoogle Scholar
  8. Japanese Gastric Cancer Association (1998) Japanese classification of gastric carcinoma. 2nd English Edition. Gastric Cancer 1:10–24CrossRefPubMedGoogle Scholar
  9. Jones JL, Glynn P, Walker RA (1999) Expression of MMP-2 and MMP-9, their inhibitors, and the activator MT1-MMP in primary breast carcinomas. J Pathol 189:161–168CrossRefPubMedGoogle Scholar
  10. Kallakury BV, Karikehalli S, Haholu A, Sheehan CE, Azumi N, Ross JS (2001) Increased expression of matrix metalloproteinases 2 and 9 and tissue inhibitors of metalloproteinases 1 and 2 correlate with poor prognostic variables in renal cell carcinoma. Clin Cancer Res 7:3113–3119PubMedGoogle Scholar
  11. Kohn EC, Liotta LA (1995) Molecular insights into cancer invasion: strategies for prevention and intervention. Cancer Res 55:1856–1862PubMedGoogle Scholar
  12. Kuraoka K, Matsumura S, Hamai Y, Nakachi K, Imai K, Matsusaki K, Oue N, Ito R, Nakayama H, Yasui W (2003) A single nucleotide polymorphism in the transmenbrane domain coding region of HER-2 is associated with development and malignant phenotype of gastric cancer. Int J Cancer 107:593–596CrossRefPubMedGoogle Scholar
  13. Lauren P (1965) The two histological main types of gastric carcinoma. Diffuse and so-called intestinal type carcinoma: an attempt at histological classification. Acta Pathol Microbiol Scand 64:31–49PubMedGoogle Scholar
  14. Liotta LA, Steeg PS, Stetler-Stevenson WG (1991) Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation. Cell 64:327–336CrossRefPubMedGoogle Scholar
  15. Matsumura S, Oue N, Kitadai Y, Chayama K, Yoshida K, Yamaguchi Y, Toge T, Imai K, Nakachi K, Yasui W (2004) A single nucleotide polymorphism in the MMP-1 promoter is correlated with histological differentiation of gastric cancer. J Cancer Res Clin Oncol 130:259–265CrossRefGoogle Scholar
  16. McDonnell S, Matrisian LM (1990) Stromelysin in tumor progression and metastasis. Cancer Metastasis Rev 9:305–319PubMedGoogle Scholar
  17. Murray GI, Duncan ME, Arbuckle E, Melvin WT, Fothergill JE (1998) Matrix metalloproteinases and their inhibitors in gastric cancer. Gut 43:791–797PubMedGoogle Scholar
  18. Nakayama T, Ito M, Ohtsuru A, Naito S, Nakashima M, Fagin JA, Yamashita S, Sekine I (1996) Expression of the Ets-1 proto-oncogene in human gastric carcinoma: correlation with tumor invasion. Am J Pathol 149:1931–1939PubMedGoogle Scholar
  19. Nagase H, Woessner JF Jr (1999) Matrix metalloproteinases. J Biol Chem 274:21491–494CrossRefPubMedGoogle Scholar
  20. Oue N, Motoshita J, Yokozaki H, Hayashi K, Tahara E, Taniyama K, Matsusaki K, Yasui W (2002) Distinct promoter hypermethylation of p16INK4a, CDH1, and RAR-ß in intestinal, diffuse-adherent, and diffuse-scattered type gastric carcinomas. J Pathol 198:55–59CrossRefPubMedGoogle Scholar
  21. Oue N, Oshimo Y, Nakayama H, Ito R, Yoshida K, Matsusaki K, Yasui W (2003) DNA methylation of multiple genes in gastric carcinoma: association with histological type and CpG island methylator phenotype. Cancer Sci 94:901–905PubMedGoogle Scholar
  22. Ozalp S, Tanir HM, Yalcin OT, Kabukcuoglu S, Oner U, Uray M (2003) Prognostic value of matrix metalloproteinase-9 (gelatinase-B) expression in epithelial ovarian tumors. Eur J Gynaecol Oncol 24:417–420PubMedGoogle Scholar
  23. Park CC, Bissell MJ, Barcellos-Hoff MH (2000) The influence of the microenvironment on the malignant phenotype. Mol Med Today 6:324–329CrossRefPubMedGoogle Scholar
  24. Rao JS, Steck PA, Mohanam S, Stetler-Stevenson WG, Liotta LA, Sawaya R (1993) Elevated levels of M(r) 92,000 type IV collagenase in human brain tumors. Cancer Res 53:2208–2211PubMedGoogle Scholar
  25. Sakata K, Satoh M, Someya M, Asanuma H, Nagakura H, Oouchi A, Nakata K, Kogawa K, Koito K, Hareyama M, Himi T (2004) Expression of matrix metalloproteinase 9 is a prognostic factor in patients with non-Hodgkin lymphoma. Cancer 100:356–365CrossRefPubMedGoogle Scholar
  26. Sehgal I, Baley PA, Thompson TC (1996) Transforming growth factor beta1 stimulates contrasting responses in metastatic versus primary mouse prostate cancer-derived cell lines in vitro. Cancer Res 56:3359–3365PubMedGoogle Scholar
  27. Sobin LH, Wittekind CH (eds) (2002) TNM classification of malignant tumors, 6th edn. Wiley-Liss, New York, pp 65–68Google Scholar
  28. Tanioka Y, Yoshida T, Yagawa T, Saiki Y, Takeo S, Harada T, Okazawa T, Yanai H, Okita K (2003) Matrix metalloproteinase-7 and matrix metalloproteinase-9 are associated with unfavourable prognosis in superficial oesophageal cancer. Br J Cancer 89:2116–2121CrossRefPubMedGoogle Scholar
  29. Wroblewski LE, Pritchard DM, Carter S, Varro A (2002) Gastrin-stimulated gastric epithelial cell invasion: the role and mechanism of increased matrix metalloproteinase 9 expression. Biochem J 365:873–879PubMedGoogle Scholar
  30. Yasui W, Yokozaki H, Fujimoto J, Naka K, Kuniyasu H, Tahara E (2000) Genetic and epigenetic alterations in multistep carcinogenesis of the stomach. J Gastroenterol 35:111–115PubMedGoogle Scholar
  31. Zhang B, Ye S, Hermann SM, Eriksson P, de Maat M, Evans A, Arveiler D, Luc G, Cambien F, Hamsten A, Watkins H, Henney AM (1999) Functional polymorphism in the regulatory region of gelatinase B gene in relation to severity of coronary atherosclerosis. Circulation 99:1788–794PubMedGoogle Scholar
  32. Zhang S, Li L, Lin JY, Lin H (2003) Imbalance between expression of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in invasiveness and metastasis of human gastric carcinoma. World J Gastroenterol 9:899–904PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Shunji Matsumura
    • 1
  • Naohide Oue
    • 1
  • Hirofumi Nakayama
    • 1
  • Yasuhiko Kitadai
    • 2
  • Kazuhiro Yoshida
    • 3
  • Yoshiyuki Yamaguchi
    • 3
  • Kazue Imai
    • 4
  • Kei Nakachi
    • 4
  • Keisuke Matsusaki
    • 5
  • Kazuaki Chayama
    • 2
  • Wataru Yasui
    • 1
    Email author
  1. 1.Department of Molecular PathologyHiroshima University Graduate School of Biomedical SciencesHiroshimaJapan
  2. 2.Department of Medicine and Molecular ScienceHiroshima University Graduate School of Biomedical SciencesHiroshimaJapan
  3. 3.Department of Surgical Oncology, Research Institute for Radiation Biology and MedicineHiroshima UniversityHiroshimaJapan
  4. 4.Department of Radiobiology/Molecular EpidemiologyRadiation Effects Research FoundationHiroshimaJapan
  5. 5.Department of SurgeryHofu Institute of GastroenterologyYamaguchiJapan

Personalised recommendations