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Hypomethylation of the metastasis-associated S100A4 gene correlates with gene activation in human colon adenocarcinoma cell lines

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Abstract

The DNA methylation status of the metastasis-associated S100A4 gene in S100A4-positive and -negative human colon adenocarcinoma cell lines was examined. Northern and Western blot analyses revealed that HT-29, SW480, SW620, WiDr and Colo201 cells expressed S100A4, whereas SW837, LoVo and DLD-1 cells expressed little S100A4. Using CpG methylation-sensitive and -insensitive restriction enzymes and PCR-based methylation assay, it was found that the S100A4 gene in HT-29, SW480, SW620, WiDr and Colo201 cells, but not in SW837, LoVo and DLD-1 cells, was hypomethylated and that the hypomethylation of the second intron was correlated well with the expression of S100A4. 5-Aza-2'-deoxycytidine, an inhibitor of the eukaryotic DNA methyltransferase, induced the expression of the S100A4 gene in SW837, LoVo and DLD-1 cells, while it showed no effect on the expression of the gene in WiDr cells. These results indicate that hypomethylation of the S100A4 gene results in the expression of the gene in colon adenocarcinoma cells. © Rapid Science Ltd.

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References

  1. Celio MR, Pauls T and Schwaller B, 1996, Guidebook to the Calcium-Binding Proteins. Oxford University Press: Sambrook and Tooze.

    Google Scholar 

  2. Hilt DC and Kligman D, 1991, The S-100 protein family: a biochemical and functional overview. In: Heizman CW, ed. Novel Calcium-binding Proteins. Fundamental and Clinical Implications. Berlin: Springer-Verlag, pp. 65-103.

    Google Scholar 

  3. Schäfer BW and Heizmann CW, 1996, The S100 family of EF-hand calcium-binding proteins: functions and pathology. Trends Biochem Sci, 21, 134-40.

    PubMed  Google Scholar 

  4. Goto K, Endo H and Fujiyoshi T, 1988, Cloning of the sequences expressed abundantly in established cell lines: identification of a cDNA clone highly homologous to S-100, a calcium binding protein. J Biochem, 103, 48-53.

    PubMed  CAS  Google Scholar 

  5. Jackson-Grusby LL, Swiergiel J and Linzer DIH, 1987, A growth-related mRNA in cultured mouse cells encodes a placental calcium binding protein. Nucleic Acid Res, 15, 6677-90.

    Article  PubMed  CAS  Google Scholar 

  6. Barraclough R, Savin J, Dube SK and Rudland PS, 1987, Molecular cloning and sequence of the gene for p9Ka, a cultured myoepithelial cell protein with strong homology to S-100, a calcium-binding protein. J Mol Biol, 198, 13-20.

    Article  PubMed  CAS  Google Scholar 

  7. Strutz F, Okada H, Lo CW, et al. 1995, Identification and characterization of a fibroblast marker: FSP1. J Cell Biol, 130, 393-405.

    Article  PubMed  CAS  Google Scholar 

  8. Ebralidze A, Tulchinsky E, Grigorian M, et al. 1989, Isolation and characterization of a gene specifically expressed in different metastatic cells and whose deduced gene product has a high degree of homology to a Ca2+-binding protein family. Genes Devl, 3, 1086-93.

    Article  CAS  Google Scholar 

  9. Takenaga K, Nakamura Y, Endo H and Sakiyama S, 1994, Involvement of S100-related calcium-binding protein pEL98 (or mts1) in cell motility and tumor cell invasion. Jpn J Cancer Res, 85, 831-9.

    PubMed  CAS  Google Scholar 

  10. Davies BR, Davies MPA, Gibbs FEM, Barraclough R and Rudland PS, 1993, Induction of the metastatic phenotype by transfection of a benign rat mammary epithelial cell line with the gene for p9Ka, a rat calcium-binding protein, but not with the oncogene EJ-ras-1. Oncogene, 8, 999-1008.

    PubMed  CAS  Google Scholar 

  11. Parker C, Whittaker PA, Usmani BA, Lakshmi MS and Sherbet GV, 1994, Induction of 18A2/mts1 gene expression and its effect on metastasis and cell cycle control. DNA Cell Biol, 13, 1021-8.

    PubMed  CAS  Google Scholar 

  12. Grigorian MS, Tulchinsky EM, Zain S, et al. 1993, The mts1 gene and control of tumor metastasis. Gene, 135, 229-38.

    Article  PubMed  CAS  Google Scholar 

  13. Takenaga K, Nakamura Y and Sakiyama S, 1997, Expression of antisense RNA to S100A4 gene encoding an S100-related calcium-binding protein suppresses metastatic potential of high-metastatic Lewis lung carcinoma cells. Oncogene, 14, 331-7.

    Article  PubMed  CAS  Google Scholar 

  14. Mælandsmo GM, Hovig E, Skrede M, et al. 1996, Reversal of the in vivometastatic phenotype of human tumor cells by anti-CAPL (mts1) ribozyme. Cancer Res, 56, 5490-8.

    PubMed  Google Scholar 

  15. Ambartsumian NS, Grigorian MS, Larsen IF, et al. 1996, Metastasis of mammary carcinomas in GRS/A hybrid mice transgenic for the mts1 gene. Oncogene, 13, 1621-30.

    PubMed  CAS  Google Scholar 

  16. Davies MPA, Rudland PS, Robertson L, Parry EW, Jolicoeur P and Barraclough R, 1996, Expression of the calcium-binding protein S100A4 (p9Ka) in MMTV-neu transgenic mice induces metastasis of mammary tumors. Oncogene, 13, 1631-7.

    PubMed  CAS  Google Scholar 

  17. Schäfer BW, Wicki R, Engelkamp D, Mattei M-G and Heizmann CW, 1995, Isolation of a YAC clone covering a cluster of nine S100 genes on human chromosome 1q21: rationale for a new nomenclature of the S100 calcium-binding protein family. Genomics, 25, 638-43.

    Article  PubMed  Google Scholar 

  18. Engelkamp D, Schäfer BW, Mattei MG, Erne P and Heizmann CW, 1993, Six S100 genes are clustered on human chromosome 1q21: identification of two genes coding for the two previously unreported calcium-binding proteins S100D and S100E. Proc Natl Acad Sci USA, 90, 6547-51.

    Article  PubMed  CAS  Google Scholar 

  19. Ambartsumian N, Tarabykina S, Grigorian M, et al. 1995, Characterization of two splice variants of metastasis-associated human mts1 gene. Gene, 159, 125-30.

    Article  PubMed  CAS  Google Scholar 

  20. Tulchinsky E, Kramerov D, Ford HL, Reshetnyak E, Lukanidin E and Zain S, 1993, Characterization of a positive regulatory element in the mts1 gene. Oncogene, 8, 79-86.

    PubMed  CAS  Google Scholar 

  21. Tulchinsky EM, Georgiev GP and Lukanidin EM, 1993, Novel AP-1 binding site created by DNA-methylation. Oncogene, 12, 1737-45.

    Google Scholar 

  22. Chen D-S and Barraclough R, 1996, Regulatory elements in the first intron of the rat S100A4 (p9Ka) gene. Biochem Soc Transact, 24, 352S.

    Google Scholar 

  23. Tulchinsky E, Ford HL, Kramerov FD, et al. 1992, Transcriptional analysis of the mts1 gene with specific reference to 5´ flanking sequences. Proc Natl Acad Sci USA, 89, 9146-50.

    Article  PubMed  CAS  Google Scholar 

  24. Tulchinsky E, Grigorian M, Tkatch T, Georgiev G and Lukanidin E, 1995, Transcriptional regulation of the mts1 gene in human lymphoma cells: the role of DNA-methylation. Biochim Biophys Acta, 1261, 243-8.

    PubMed  Google Scholar 

  25. Takenaga K, Nakanishi H, Wada K, et al. 1997, Increased expression of S100A4, a metastasis-associated gene, in human colorectal adenocarcinomas. Clin Cancer Res, 3, 2309-16.

    PubMed  CAS  Google Scholar 

  26. Sambrook J, Fritsch EF and Maniatis T, 1989, Molecular Cloning: a laboratory manual, 2nd edn. New York: Cold Spring Harbor Laboratory.

    Google Scholar 

  27. Takenaga K, Nakamura Y, Sakiyama S, Hasegawa Y, Sato K and Endo H, 1994, Binding of pEL98 protein, an S100-related calcium-binding protein, to nonmuscle tropomyosin. J Cell Biol, 124, 757-68.

    Article  PubMed  CAS  Google Scholar 

  28. Laemmli UK, 1970, Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-5.

    Article  PubMed  CAS  Google Scholar 

  29. Towbin H, Staehelin T and Gordon J, 1979, Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA, 76, 4350-4.

    Article  PubMed  CAS  Google Scholar 

  30. Bradford MM, 1976, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem, 72, 248-54.

    Article  PubMed  CAS  Google Scholar 

  31. Schumutte C, Yang AS, Nguyen TT, Beart RW and Jones PA, 1996, Mechanisms for the involvement of DNA methylation in colon carcinogenesis. Cancer Res, 56, 2375-81.

    Google Scholar 

  32. Yamamoto T, Gotoh M, Kitajima M and Hirohashi S, 1993, Thymosin β-4 expression is correlated with metastatic capacity of colorectal carcinomas. Biochem Biophys Res Commun, 193, 706-10.

    Article  PubMed  CAS  Google Scholar 

  33. Vermeulen SJ, Bruyneel EA, Bracke ME, et al. 1995, Transition from the noninvasive to the invasive phenotype and loss of α-catenin in human colon cancer cells. Cancer Res, 55, 4722-8.

    PubMed  CAS  Google Scholar 

  34. Aznavoorian S, Liotta LA and Kupchik HZ, 1990, Characteristics of invasive and noninvasive human colorectal adenocarcinoma cells. J Natl Cancer Inst, 82, 1485-92.

    Article  PubMed  CAS  Google Scholar 

  35. Zirvi KA, Keogh JP, Slomiany A and Slomiany BL, 1991, Transglutaminase activity in human colorectal carcinomas of differing metastatic potential. Cancer Lett, 60, 85-92.

    Article  PubMed  CAS  Google Scholar 

  36. Kuo T-H, Kubota T, Watanabe M, et al. 1995, Liver colonization competence governs colon cancer metastasis. Proc Natl Acad Sci USA, 92, 12085-9.

    Article  PubMed  CAS  Google Scholar 

  37. Herzberg F, Schöning M, Schirner M, Topp M, Thiel E and Kreuser E-D, 1996, IL-4 and TNF-? induce changes in integrin expression and adhesive properties and decrease the lung-colonizing potential of HT-29 colon carcinoma cells. Clin Exp Metastasis, 14, 165-75.

    Article  PubMed  CAS  Google Scholar 

  38. Jones PA and Buckley JD, 1990, The role of DNA methylation in cancer. Adv Cancer Res, 54, 1-23.

    Article  PubMed  CAS  Google Scholar 

  39. Issa J-PJ, Vertino PM, Wu J, et al. 1993, Increased cytosine DNA-methyltransferase activity during colon cancer progression. J Natl Cancer Inst, 85, 1235-40.

    Article  PubMed  CAS  Google Scholar 

  40. Szyf M, Theberge J and Bozoric V, 1995, Ras induces a general DNA demethylation activity in mouse embryonal P19 cells. J Biol Chem, 270, 12690-6.

    Article  PubMed  CAS  Google Scholar 

  41. Takenaga K, Nakamura Y and Sakiyama S, 1994, Cellular localization of pEL98 protein, an S100-related calcium binding protein, in fibroblasts and its tissue distribution analyzed by monoclonal antibodies. Cell Struc Funct, 19, 133-41.

    Article  CAS  Google Scholar 

  42. Litepro RG and Kerbel RS, 1987, Reduced levels of DNA 5-methylcytosine in metastatic variants of the human melanoma cell line MeWo. Cancer Res, 47, 2264-7.

    Google Scholar 

  43. Trainer DL, Kline T, Mallon F, Greig R and Poste G, 1985, Effect of 5-azacytidine on DNA methylation and the malignant properties of B16 melanoma cells. Cancer Res, 45, 6124-30.

    PubMed  CAS  Google Scholar 

  44. Habets GG, van der Kammen RA, Scholtes EH and Collard JG, 1990, Induction of invasive and metastatic potential in mouse T-lymphoma cells (BW5147) by treatment with 5-azacytidine. Clin Exp Metastasis, 8, 567-77.

    Article  PubMed  CAS  Google Scholar 

  45. Ormerod EJ, Everett CA and Hart IR, 1986, Enhanced experimental capacity of a human tumor line following treatment with 5-azacytidine. Cancer Res, 46, 884-90.

    PubMed  CAS  Google Scholar 

  46. Yoshiura K, Kanai Y, Ochiai A, Shimoyama Y, Sugimura T and Hirohashi S, 1995, Silencing of the E-cadherin invasion-suppressor gene by CpG methylation in human carcinomas. Proc Natl Acad Sci USA, 92, 7416-9.

    Article  PubMed  CAS  Google Scholar 

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Nakamura, N., Takenaga, K. Hypomethylation of the metastasis-associated S100A4 gene correlates with gene activation in human colon adenocarcinoma cell lines. Clin Exp Metastasis 16, 471–479 (1998). https://doi.org/10.1023/A:1006589626307

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