Identification by differential display of a protein phosphatase-2A regulatory subunit preferentially expressed in malignant melanoma cells

Int J Cancer. 1999 Aug 27;82(5):709-13. doi: 10.1002/(sici)1097-0215(19990827)82:5<709::aid-ijc14>3.0.co;2-r.

Abstract

We described the occurrence of 4 transcripts differentially displayed between syngeneic murine B16F10 (metastatic melanoma) and Melan-a (immortalised melanocytes) cell lines. We now report that one such transcript, which is B16F10-specific, represents a protein phosphatase-2A B' regulatory subunit. No expression of this transcript was detected in the weakly metastatic B16F1 by Northern blotting. Moreover, the transcript was not expressed by spontaneously immortalised, non-tumorigenic, melanocytes (Melan-Ab and Melan-a2), nor was it expressed by ras-transformed, tumourigenic melanocytes (Melan-Ab-LTR-ras). Cloning of the 5'-end region of this transcript (termed band 8A) from B16F10 cells revealed an intracisternal A-particle insertion, including the long terminal repeat region, which could account for the observed high expression in B16F10 cells. Single cell clones of B16F10 manifested an experimental metastasis capacity, which correlated with band 8A expression with the lowest expressors being least metastatic. The human homologue of the B' regulatory subunit, B56gamma, is expressed preferentially at the mRNA level in human melanoma cell lines compared with normal epidermal melanocytes. In situ hybridisation studies on human clinical samples detected high expression of this gene in a number of malignant melanomas. Our results imply strongly that this protein phosphatase-2A regulatory subunit may have a role in melanoma tumour progression.

MeSH terms

  • Animals
  • Base Sequence
  • Blotting, Northern
  • Blotting, Western
  • Cell Line
  • Cloning, Molecular
  • Humans
  • In Situ Hybridization
  • Melanoma, Experimental / enzymology*
  • Melanoma, Experimental / pathology
  • Mice
  • Molecular Sequence Data
  • Neoplasm Metastasis
  • Phosphoprotein Phosphatases / biosynthesis
  • Phosphoprotein Phosphatases / isolation & purification*
  • Protein Phosphatase 2
  • Tumor Cells, Cultured

Substances

  • Phosphoprotein Phosphatases
  • Protein Phosphatase 2