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      Gene expression patterns in melanocytic cells: candidate markers for early stage and malignant transformation.

      The Journal of Pathology
      metabolism, Blotting, Northern, Cell Culture Techniques, Cell Transformation, Neoplastic, genetics, DNA, Complementary, DNA, Neoplasm, Gene Expression, Humans, Melanocytes, Melanoma, secondary, Neoplasm Proteins, Nevus, Pigmented, Polymerase Chain Reaction, methods, Skin Neoplasms, Tumor Markers, Biological, rab5 GTP-Binding Proteins

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          Abstract

          Different stages of differentiation of human melanocytic cells, such as normal melanocytes, naevus and melanoma cells, reflect distinct gene expression patterns. A PCR-based subtractive hybridization and display method was applied to identify genes that are differentially expressed in melanocytic cells in relation to early stage and malignant transformation. This resulted in the identification of a number of candidate cDNAs differentially expressed among melanocytes, naevus cells, and (non)-metastatic melanoma cells. Out of this collection of cDNAs, 16 clones were screened that comprised 12 novel genes, one previously identified expressed sequence tag related to vesicular trafficking (Ras-related protein Rab5b). The other three were also known genes that were either related to cell motility (beta-tubulin), pre-mRNA splicing (small nuclear protein U1A), or of unknown function (the human TI227-H gene). The differential expression patterns of Rab5b and two novel gene fragments (pCMa1, pCMn2) were further assessed in melanocytic cells. pCMa1 was expressed more in metastatic melanoma than in primary melanoma cells. In contrast, pCMn2 was expressed in both non-metastatic and metastatic melanoma cells, but was not detectable in either normal melanocytes or naevus cells. The Ras-related protein Rab5b showed lower levels of expression in highly metastatic than in other melanoma cells. These three cDNAs may therefore be involved in the early stage and malignant transformation of melanocytes. Copyright 2001 John Wiley & Sons, Ltd.

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