Abstract

Expression of protein kinase C (PKC) genes (alpha, beta, gamma and epsilon) was measured in cultured normal human neonatal melanocytes and metastatic melanoma cell strains. Three of the PKC isotypes (alpha, beta and epsilon) were constitutively expressed in neonatal melanocytes. Protein kinase C beta RNA transcripts were induced in neonatal melanocytes cultivated in medium with serum and 12-O-tetradecanoylphorbol-13-acetate (TPA). In contrast, PKC alpha and epsilon RNA transcripts were detected in melanocytes cultivated in medium without serum and TPA, but were repressed in melanocytes cultivated in medium with serum and TPA. Only PKC alpha and epsilon RNA transcripts were detected in the melanoma cell strains and the PKC RNA transcript expression levels varied among the five metastatic melanomas. In four metastatic melanoma cell strains, PKC alpha and epsilon RNA transcript expression levels were repressed by serum, but in one melanoma cell strain, PKC alpha and epsilon RNA transcript expression levels were induced by serum. Protein kinase C gamma RNA transcripts were not detected in either the melanocytes or melanoma cell strains. These data suggest an alteration of PKC isotype gene expression in the progression of primary melanocytes to metastatic melanoma. The absence of the PKC beta RNA transcripts and altered expression of PKC alpha and epsilon isotypes in particular may be a feature in the transformation of human primary melanocytes.

Highlights

  • Within the past decade, the cultivation of human primary melanocytes in vitro has become possible

  • Expression of the protein kinase C (PKC) a 4.3 kb RNA transcript was repressed in three melanoma strains [c8M6a (59%), c81^6c (72%) and c81-61 x (49%)] when the cells were cultivated in medium with serum compared to medium without serum

  • We have investigated the expression of PKC (a, /3, y and e) RNA transcripts in human primary neonatal melanocytes and metastatic melanoma cell strains

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Summary

Introduction

The cultivation of human primary melanocytes in vitro has become possible. The requirements for growth were phorbol esters (e.g. 12-

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