Galectins pp 355-369 | Cite as

Examination of the Regulation of Galectin-3 Expression in Cancer

  • Hafiz AhmedEmail author
  • Gargi Bandyopadhyaya
Part of the Methods in Molecular Biology book series (MIMB, volume 1207)


Galectin-3, a member of a β-galactoside-binding protein family, is involved in normal growth development as well as cancer progression and metastasis, but the detailed mechanisms of its functions or its transcriptional regulations are not well understood. Besides, several regulatory elements such as GC box, CRE motif, AP-1 site, and NF-κB sites, the promoter of galectin-3 gene (LGALS3) contains several CpG islands that can be methylated during tumorigenesis of prostate leading to the gene silencing. Here we describe protocols for identification of galectin-3 DNA methylation, suppression of DNA methyltransferases to reactivate galectin-3 expression, and development of methylation-specific polymerase chain reaction (MS-PCR) to assess galectin-3 expression in various biological specimens such as tissue, serum, and urine samples.

Key words

Galectin-3 Gene regulation Gene silencing DNA methylation Prostate cancer 



The work carried out by us was supported by the US Army Medical Research and Materiel Command grant W81XWH-07-1-0565, a start-up fund from the University of Maryland School of Medicine, and the National Institute of Health Grants CA133935 and CA141970 to H.A.


  1. 1.
    Nakahara S, Raz A (2007) Regulation of cancer-related gene expression by galectin-3 and the molecular mechanism of its nuclear import pathway. Cancer Metastasis Rev 26:605–610CrossRefPubMedCentralPubMedGoogle Scholar
  2. 2.
    Rabinovich GA, Liu FT, Hirashima M et al (2007) An emerging role for galectins in tuning the immune response: lessons from experimental models of inflammatory disease, autoimmunity and cancer. Scand J Immunol 66:143–158CrossRefPubMedGoogle Scholar
  3. 3.
    Newlaczyl AU, Yu LG (2011) Galectin-3–a jack-of-all-trades in cancer. Cancer Lett 313:123–128CrossRefPubMedGoogle Scholar
  4. 4.
    Ahmed H, Guha P, Kaptan E et al (2011) Galectin-3: a potential target for cancer prevention. Trend Carbohydr Res 3:13–22Google Scholar
  5. 5.
    Braeuer RR, Shoshan E, Kamiya T et al (2012) The sweet and bitter sides of galectins in melanoma progression. Pigment Cell Melanoma Res 25:592–601CrossRefPubMedGoogle Scholar
  6. 6.
    Danguy A, Camby I, Kiss R (2002) Galectins and cancer. Biochim Biophys Acta 1572:285–293CrossRefPubMedGoogle Scholar
  7. 7.
    Califice S, Castronovo V, Van Den Brûle F (2004) Galectin-3 and cancer. Int J Oncol 25:983–992PubMedGoogle Scholar
  8. 8.
    Hsu DK, Dowling CA, Jeng KC et al (1999) Galectin-3 expression is induced in cirrhotic liver and hepatocellular carcinoma. Int J Cancer 81:519–526CrossRefPubMedGoogle Scholar
  9. 9.
    Miyazaki J, Hokari R, Kato S et al (2002) Increased expression of galectin-3 in primary gastric cancer and the metastatic lymph nodes. Oncol Rep 9:1307–1312PubMedGoogle Scholar
  10. 10.
    Yoshimura A, Gemma A, Hosoya Y et al (2003) Increased expression of the LGALS3 (galectin 3) gene in human non-small-cell lung cancer. Genes Chromosomes Cancer 37:159–164CrossRefPubMedGoogle Scholar
  11. 11.
    Sakaki M, Oka N, Nakanishi R et al (2008) Serum level of galectin-3 in human bladder cancer. J Med Invest 55:127–132CrossRefPubMedGoogle Scholar
  12. 12.
    Saussez S, Glinoer D, Chantrain G, Pattou F et al (2008) Serum galectin-1 and galectin-3 levels in benign and malignant nodular thyroid disease. Thyroid 18:705–712CrossRefPubMedGoogle Scholar
  13. 13.
    Saussez S, Decaestecker C, Mahillon V et al (2008) Galectin-3 upregulation during tumor progression in head and neck cancer. Laryngoscope 118:1583–1590CrossRefPubMedGoogle Scholar
  14. 14.
    Pacis RA, Pilat MJ, Pienta KJ et al (2000) Decreased galectin-3 expression in prostate cancer. Prostate 44:118–123CrossRefPubMedGoogle Scholar
  15. 15.
    Merseburger AS, Kramer MW, Hennenlotter J et al (2008) Involvement of decreased galectin-3 expression in the pathogenesis and progression of prostate cancer. Prostate 68:72–77CrossRefPubMedGoogle Scholar
  16. 16.
    Ahmed H, Cappello F, Rodolico V et al (2009) Evidence of heavy methylation in the galectin-3 promoter in early stages of prostate adenocarcinoma: development and validation of a methylated marker for early diagnosis of prostate cancer. Transl Oncol 2:146–156CrossRefPubMedCentralPubMedGoogle Scholar
  17. 17.
    Merseburger AS, Kramer MW, Hennenlotter J et al (2008) Loss of galectin-3 expression correlates with clear cell renal carcinoma progression and reduced survival. World J Urol 26:637–642CrossRefPubMedGoogle Scholar
  18. 18.
    Ruebel KH, Jin L, Qian X et al (2005) Effects of DNA methylation on galectin-3 expression in pituitary tumors. Cancer Res 65:1136–1140CrossRefPubMedGoogle Scholar
  19. 19.
    Honjo Y, Inohara H, Akahani S et al (2000) Expression of cytoplasmic galectin-3 as a prognostic marker in tongue carcinoma. Clin Cancer Res 6:4635–4640PubMedGoogle Scholar
  20. 20.
    Dumic J, Dabelic S, Flögel M (2006) Galectin-3: an open-ended story. Biochem Biophys Acta 1760:616–635CrossRefPubMedGoogle Scholar
  21. 21.
    Kadrofske MM, Openo KP, Wang JL (1998) The human LGALS3 (galectin-3) gene: determination of the gene structure and functional characterization of the promoter. Arch Biochem Biophys 349:7–20CrossRefPubMedGoogle Scholar
  22. 22.
    Fogel S, Guittaut M, Legrand A et al (1999) The tat protein of HIV-1 induces galectin-3 expression. Glycobiology 9:383–387CrossRefPubMedGoogle Scholar
  23. 23.
    Hsu DK, Hammes SR, Kuwabara I et al (1996) Human T lymphotropic virus-I infection of human T lymphocytes induces expression of the beta-galactoside-binding lectin, galectin-3. Am J Pathol 148:1661–1670PubMedCentralPubMedGoogle Scholar
  24. 24.
    Dumic J, Lauc G, Flogel M (2000) Expression of galectin-3 in cells exposed to stress-roles of jun and NF-kappaB. Cell Physiol Biochem 10:149–158CrossRefPubMedGoogle Scholar
  25. 25.
    Stock M, Schafer H, Stricker S et al (2003) Expression of galectin-3 in skeletal tissues is controlled by Runx2. J Biol Chem 278:17360–17367CrossRefPubMedGoogle Scholar
  26. 26.
    Costessi A, Pines A, D’Andrea P et al (2005) Extracellular nucleotides activate Runx2 in the osteoblast-like HOBIT cell line: a possible molecular link between mechanical stress and osteoblasts’ response. Bone 36:418–432CrossRefPubMedGoogle Scholar
  27. 27.
    Nakahara S, Oka N, Raz A (2005) On the role of galectin-3 in cancer apoptosis. Apoptosis 10:267–275CrossRefPubMedGoogle Scholar
  28. 28.
    Ahmed H, Banerjee PB, Vasta GR (2007) Differential expression of galectins in normal, benign and malignant prostate epithelial cells: silencing of galectin-3 expression in prostate cancer by its promoter methylation. Biochem Biophys Res Commun 358:241–246CrossRefPubMedGoogle Scholar
  29. 29.
    Ahmed H (2010) Promoter methylation in prostate cancer and its application for the early detection of prostate cancer using serum and urine samples. Biomark Cancer 2010:17–33CrossRefPubMedGoogle Scholar
  30. 30.
    McKenna ES, Roberts CW (2009) Epigenetics and cancer without genomic instability. Cell Cycle 8:23–26CrossRefPubMedGoogle Scholar
  31. 31.
    Bestor TH (1992) Activation of mammalian DNA methyltransferase by cleavage of a Zn binding regulatory domain. EMBO J 11:2611–2617PubMedCentralPubMedGoogle Scholar
  32. 32.
    Robert MF, Morin S, Beaulieu N et al (2003) DNMT1 is required to maintain CpG methylation and aberrant gene silencing in human cancer cells. Nat Genet 33:61–65CrossRefPubMedGoogle Scholar
  33. 33.
    El-Osta A (2003) DNMT cooperativity–the developing links between methylation, chromatin structure and cancer. Bioessays 25:1071–1084CrossRefPubMedGoogle Scholar
  34. 34.
    Benbrahim-Tallaa L, Waterland RA, Dill AL et al (2007) Tumor suppressor gene inactivation during cadmium-induced malignant transformation of human prostate cells correlates with overexpression of de novo DNA methyltransferase. Environ Health Perspect 115:1454–1459PubMedCentralPubMedGoogle Scholar
  35. 35.
    Roll JD, Rivenbark AG, Jones WD et al (2008) DNMT3b overexpression contributes to a hypermethylator phenotype in human breast cancer cell lines. Mol Cancer 7:15CrossRefPubMedCentralPubMedGoogle Scholar
  36. 36.
    Tate PH, Bird AP (1993) Effects of DNA methylation on DNA-binding proteins and gene expression. Curr Opin Genet Dev 3:226–231CrossRefPubMedGoogle Scholar
  37. 37.
    Hendrich B, Bird A (1998) Identification and characterization of a family of mammalian methyl-CpG binding proteins. Mol Cell Biol 18:6538–6547PubMedCentralPubMedGoogle Scholar
  38. 38.
    Sansom OJ, Maddison K, Clarke AR (2007) Mechanisms of disease: methyl-binding domain proteins as potential therapeutic targets in cancer. Nat Clin Pract Oncol 4:305–315CrossRefPubMedGoogle Scholar
  39. 39.
    Zhang Y, Ng HH, Erdjument-Bromage H et al (1999) Analysis of the NuRD subunits reveals a histone deacetylase core complex and a connection with DNA methylation. Genes Dev 13:1924–1935CrossRefPubMedCentralPubMedGoogle Scholar
  40. 40.
    Nan X, Ng HH, Johnson CA et al (1998) Transcriptional repression by the methyl-CpG-binding protein MeCP2 involves a histone deacetylase complex. Nature 393:386–389CrossRefPubMedGoogle Scholar
  41. 41.
    Tyler JK, Kadonaga JT (1999) The “dark side” of chromatin remodeling: repressive effects on transcription. Cell 99:443–446CrossRefPubMedGoogle Scholar
  42. 42.
    Li LC, Dahiya R (2002) MethPrimer: designing primers for methylation PCRs. Bioinformatics 18:1427–1431CrossRefPubMedGoogle Scholar
  43. 43.
    Ahmed H (2012) Methylated DNA as promising marker for early diagnosis of cancer. J Bioanal Biomed 4:e108Google Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Biochemistry and Molecular Biology, Institute of Marine and Environmental Technology, Greenebaum Cancer CenterUniversity of Maryland School of MedicineBaltimoreUSA
  2. 2.Department of Biochemistry and Molecular Biology, Institute of Marine and Environmental TechnologyUniversity of Maryland School of MedicineBaltimoreUSA

Personalised recommendations