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. 1995 Aug 1;92(16):7416–7419. doi: 10.1073/pnas.92.16.7416

Silencing of the E-cadherin invasion-suppressor gene by CpG methylation in human carcinomas.

K Yoshiura 1, Y Kanai 1, A Ochiai 1, Y Shimoyama 1, T Sugimura 1, S Hirohashi 1
PMCID: PMC41350  PMID: 7543680

Abstract

E-Cadherin, a cell adhesion molecule, which plays a key role in maintaining the epithelial phenotype, is regarded as an invasion-suppressor gene in light of accumulating evidence from in vitro experiments and clinical observations. In an attempt to clarify the mechanism responsible for inactivation of this gene in carcinomas, we investigated the methylation state around the promoter region by digestion of DNA with the methylation-sensitive restriction enzyme Hpa II, as CpG methylation of the promoter has been postulated to be a mechanism of transcriptional inactivation of some genes. We found that E-cadherin expression-negative carcinoma cell lines were accompanied by the hypermethylation state, whereas E-cadherin-positive cell lines were not. Furthermore, treatment of E-cadherin-negative carcinoma cells with the demethylating agent 5-azacytidine resulted in reexpression of the gene and reversion of scattered spindle-shaped cells to cells with epithelial morphology. These results suggest that hypermethylation around the promoter may be a mechanism of E-cadherin inactivation in human carcinomas and that treatment of E-cadherin-inactivated cells with a demethylating agent may cause gene expression reversion leading to epithelial morphogenesis with acquisition of the homophilic cell-cell adhesive property.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Antequera F., Boyes J., Bird A. High levels of de novo methylation and altered chromatin structure at CpG islands in cell lines. Cell. 1990 Aug 10;62(3):503–514. doi: 10.1016/0092-8674(90)90015-7. [DOI] [PubMed] [Google Scholar]
  2. Baylin S. B., Makos M., Wu J. J., Yen R. W., de Bustros A., Vertino P., Nelkin B. D. Abnormal patterns of DNA methylation in human neoplasia: potential consequences for tumor progression. Cancer Cells. 1991 Oct;3(10):383–390. [PubMed] [Google Scholar]
  3. Becker K. F., Atkinson M. J., Reich U., Becker I., Nekarda H., Siewert J. R., Höfler H. E-cadherin gene mutations provide clues to diffuse type gastric carcinomas. Cancer Res. 1994 Jul 15;54(14):3845–3852. [PubMed] [Google Scholar]
  4. Behrens J., Löwrick O., Klein-Hitpass L., Birchmeier W. The E-cadherin promoter: functional analysis of a G.C-rich region and an epithelial cell-specific palindromic regulatory element. Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11495–11499. doi: 10.1073/pnas.88.24.11495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Behrens J., Mareel M. M., Van Roy F. M., Birchmeier W. Dissecting tumor cell invasion: epithelial cells acquire invasive properties after the loss of uvomorulin-mediated cell-cell adhesion. J Cell Biol. 1989 Jun;108(6):2435–2447. doi: 10.1083/jcb.108.6.2435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bird A. P. CpG-rich islands and the function of DNA methylation. Nature. 1986 May 15;321(6067):209–213. doi: 10.1038/321209a0. [DOI] [PubMed] [Google Scholar]
  7. Bird A. The essentials of DNA methylation. Cell. 1992 Jul 10;70(1):5–8. doi: 10.1016/0092-8674(92)90526-i. [DOI] [PubMed] [Google Scholar]
  8. Bussemakers M. J., Giroldi L. A., van Bokhoven A., Schalken J. A. Transcriptional regulation of the human E-cadherin gene in human prostate cancer cell lines: characterization of the human E-cadherin gene promoter. Biochem Biophys Res Commun. 1994 Sep 15;203(2):1284–1290. doi: 10.1006/bbrc.1994.2321. [DOI] [PubMed] [Google Scholar]
  9. D'souza B., Taylor-Papadimitriou J. Overexpression of ERBB2 in human mammary epithelial cells signals inhibition of transcription of the E-cadherin gene. Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):7202–7206. doi: 10.1073/pnas.91.15.7202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Frixen U. H., Behrens J., Sachs M., Eberle G., Voss B., Warda A., Löchner D., Birchmeier W. E-cadherin-mediated cell-cell adhesion prevents invasiveness of human carcinoma cells. J Cell Biol. 1991 Apr;113(1):173–185. doi: 10.1083/jcb.113.1.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hergersberg M. Biological aspects of cytosine methylation in eukaryotic cells. Experientia. 1991 Dec 1;47(11-12):1171–1185. doi: 10.1007/BF01918381. [DOI] [PubMed] [Google Scholar]
  12. Herman J. G., Latif F., Weng Y., Lerman M. I., Zbar B., Liu S., Samid D., Duan D. S., Gnarra J. R., Linehan W. M. Silencing of the VHL tumor-suppressor gene by DNA methylation in renal carcinoma. Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):9700–9704. doi: 10.1073/pnas.91.21.9700. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Issa J. P., Ottaviano Y. L., Celano P., Hamilton S. R., Davidson N. E., Baylin S. B. Methylation of the oestrogen receptor CpG island links ageing and neoplasia in human colon. Nat Genet. 1994 Aug;7(4):536–540. doi: 10.1038/ng0894-536. [DOI] [PubMed] [Google Scholar]
  14. Jones P. A., Taylor S. M. Hemimethylated duplex DNAs prepared from 5-azacytidine-treated cells. Nucleic Acids Res. 1981 Jun 25;9(12):2933–2947. doi: 10.1093/nar/9.12.2933. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jones P. A., Wolkowicz M. J., Rideout W. M., 3rd, Gonzales F. A., Marziasz C. M., Coetzee G. A., Tapscott S. J. De novo methylation of the MyoD1 CpG island during the establishment of immortal cell lines. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6117–6121. doi: 10.1073/pnas.87.16.6117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kanai Y., Oda T., Tsuda H., Ochiai A., Hirohashi S. Point mutation of the E-cadherin gene in invasive lobular carcinoma of the breast. Jpn J Cancer Res. 1994 Oct;85(10):1035–1039. doi: 10.1111/j.1349-7006.1994.tb02902.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Keshet I., Yisraeli J., Cedar H. Effect of regional DNA methylation on gene expression. Proc Natl Acad Sci U S A. 1985 May;82(9):2560–2564. doi: 10.1073/pnas.82.9.2560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lee W. H., Morton R. A., Epstein J. I., Brooks J. D., Campbell P. A., Bova G. S., Hsieh W. S., Isaacs W. B., Nelson W. G. Cytidine methylation of regulatory sequences near the pi-class glutathione S-transferase gene accompanies human prostatic carcinogenesis. Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11733–11737. doi: 10.1073/pnas.91.24.11733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Li E., Beard C., Jaenisch R. Role for DNA methylation in genomic imprinting. Nature. 1993 Nov 25;366(6453):362–365. doi: 10.1038/366362a0. [DOI] [PubMed] [Google Scholar]
  20. Makos M., Nelkin B. D., Lerman M. I., Latif F., Zbar B., Baylin S. B. Distinct hypermethylation patterns occur at altered chromosome loci in human lung and colon cancer. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1929–1933. doi: 10.1073/pnas.89.5.1929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mayer B., Johnson J. P., Leitl F., Jauch K. W., Heiss M. M., Schildberg F. W., Birchmeier W., Funke I. E-cadherin expression in primary and metastatic gastric cancer: down-regulation correlates with cellular dedifferentiation and glandular disintegration. Cancer Res. 1993 Apr 1;53(7):1690–1695. [PubMed] [Google Scholar]
  22. Oda T., Kanai Y., Oyama T., Yoshiura K., Shimoyama Y., Birchmeier W., Sugimura T., Hirohashi S. E-cadherin gene mutations in human gastric carcinoma cell lines. Proc Natl Acad Sci U S A. 1994 Mar 1;91(5):1858–1862. doi: 10.1073/pnas.91.5.1858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ohtani-Fujita N., Fujita T., Aoike A., Osifchin N. E., Robbins P. D., Sakai T. CpG methylation inactivates the promoter activity of the human retinoblastoma tumor-suppressor gene. Oncogene. 1993 Apr;8(4):1063–1067. [PubMed] [Google Scholar]
  24. Oka H., Shiozaki H., Kobayashi K., Inoue M., Tahara H., Kobayashi T., Takatsuka Y., Matsuyoshi N., Hirano S., Takeichi M. Expression of E-cadherin cell adhesion molecules in human breast cancer tissues and its relationship to metastasis. Cancer Res. 1993 Apr 1;53(7):1696–1701. [PubMed] [Google Scholar]
  25. Ottaviano Y. L., Issa J. P., Parl F. F., Smith H. S., Baylin S. B., Davidson N. E. Methylation of the estrogen receptor gene CpG island marks loss of estrogen receptor expression in human breast cancer cells. Cancer Res. 1994 May 15;54(10):2552–2555. [PubMed] [Google Scholar]
  26. Risinger J. I., Berchuck A., Kohler M. F., Boyd J. Mutations of the E-cadherin gene in human gynecologic cancers. Nat Genet. 1994 May;7(1):98–102. doi: 10.1038/ng0594-98. [DOI] [PubMed] [Google Scholar]
  27. Shimoyama Y., Hirohashi S. Cadherin intercellular adhesion molecule in hepatocellular carcinomas: loss of E-cadherin expression in an undifferentiated carcinoma. Cancer Lett. 1991 May 1;57(2):131–135. doi: 10.1016/0304-3835(91)90206-w. [DOI] [PubMed] [Google Scholar]
  28. Shimoyama Y., Hirohashi S. Expression of E- and P-cadherin in gastric carcinomas. Cancer Res. 1991 Apr 15;51(8):2185–2192. [PubMed] [Google Scholar]
  29. Shimoyama Y., Hirohashi S., Hirano S., Noguchi M., Shimosato Y., Takeichi M., Abe O. Cadherin cell-adhesion molecules in human epithelial tissues and carcinomas. Cancer Res. 1989 Apr 15;49(8):2128–2133. [PubMed] [Google Scholar]
  30. Shiozaki H., Tahara H., Oka H., Miyata M., Kobayashi K., Tamura S., Iihara K., Doki Y., Hirano S., Takeichi M. Expression of immunoreactive E-cadherin adhesion molecules in human cancers. Am J Pathol. 1991 Jul;139(1):17–23. [PMC free article] [PubMed] [Google Scholar]
  31. Sommers C. L., Thompson E. W., Torri J. A., Kemler R., Gelmann E. P., Byers S. W. Cell adhesion molecule uvomorulin expression in human breast cancer cell lines: relationship to morphology and invasive capacities. Cell Growth Differ. 1991 Aug;2(8):365–372. [PubMed] [Google Scholar]
  32. Sorkin B. C., Jones F. S., Cunningham B. A., Edelman G. M. Identification of the promoter and a transcriptional enhancer of the gene encoding L-CAM, a calcium-dependent cell adhesion molecule. Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11356–11360. doi: 10.1073/pnas.90.23.11356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Takeichi M. Cadherin cell adhesion receptors as a morphogenetic regulator. Science. 1991 Mar 22;251(5000):1451–1455. doi: 10.1126/science.2006419. [DOI] [PubMed] [Google Scholar]
  34. Vleminckx K., Vakaet L., Jr, Mareel M., Fiers W., van Roy F. Genetic manipulation of E-cadherin expression by epithelial tumor cells reveals an invasion suppressor role. Cell. 1991 Jul 12;66(1):107–119. doi: 10.1016/0092-8674(91)90143-m. [DOI] [PubMed] [Google Scholar]
  35. Zion M., Ben-Yehuda D., Avraham A., Cohen O., Wetzler M., Melloul D., Ben-Neriah Y. Progressive de novo DNA methylation at the bcr-abl locus in the course of chronic myelogenous leukemia. Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10722–10726. doi: 10.1073/pnas.91.22.10722. [DOI] [PMC free article] [PubMed] [Google Scholar]

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