Suppression subtractive hybridisation to isolate differentially expressed genes involved in invasiveness of melanoma cell line cultured under different conditions

Int J Oncol. 2002 Mar;20(3):501-8.


We have previously analysed the invasion capacity of different melanoma cell lines in the three-dimensional dermal equivalent. The melanoma cell line M4Beu acquired invasive behaviour upon changing its cultivation conditions before the seeding on top of the collagen lattice from single cell suspension to spheroid. Based on this phenomenon SSH was used to search for the genes related to the invasive phenotype of melanoma cells. From differentially expressed clones we focused on four: fibronectin, RhoA, COXII, and H-ras-like protein. By RT-PCR the expression of these genes were tested in different populations (monolayer, spheroids on dermal equivalent) of melanoma cell line M4Beu and three additional melanoma cell lines. The expression of fibronectin was also examined by immunohistochemistry staining of co-culture spheroids-dermal equivalent.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular
  • Cyclooxygenase 2
  • DNA, Complementary / metabolism
  • Fibronectins / biosynthesis
  • Fibronectins / genetics
  • Genetic Techniques*
  • Humans
  • Immunohistochemistry
  • Isoenzymes / biosynthesis
  • Isoenzymes / genetics
  • Melanoma / genetics*
  • Membrane Proteins
  • Neoplasm Invasiveness
  • Nucleic Acid Hybridization*
  • Polymerase Chain Reaction
  • Prostaglandin-Endoperoxide Synthases / biosynthesis
  • Prostaglandin-Endoperoxide Synthases / genetics
  • Proto-Oncogene Proteins p21(ras) / biosynthesis
  • Proto-Oncogene Proteins p21(ras) / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Tumor Cells, Cultured
  • rhoA GTP-Binding Protein / biosynthesis
  • rhoA GTP-Binding Protein / genetics


  • DNA, Complementary
  • Fibronectins
  • Isoenzymes
  • Membrane Proteins
  • Cyclooxygenase 2
  • PTGS2 protein, human
  • Prostaglandin-Endoperoxide Synthases
  • HRAS protein, human
  • Proto-Oncogene Proteins p21(ras)
  • rhoA GTP-Binding Protein