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Expression and Up-regulation of alternatively spliced transcripts of melastatin, a melanoma metastasis-related gene, in human melanoma cells.

Loss of expression of a novel suppressor of metastasis, melastatin (MLSN1), has recently been reported to correlate with metastatic potential of melanoma cells. Using differential display analysis, we identified MLSN1 among genes overexpressed in pigmented metastatic human melanoma cells treated with the differentiation inducer hexamethylene bisacetamide (HMBA). In this study, we show that multiple short transcripts of MLSN1 are present in melanocytes and pigmented metastatic melanoma cell lines while the full-length 5. 4-kb mRNA is detectable only in melanocytes. Treatment of pigmented melanoma cells with the differentiation-inducing agent, HMBA, results in up-regulation of the 5.4-kb MLSN1 mRNA as well as short RNAs. Analysis of a panel of nonpigmented primary and metastatic melanoma cell lines showed weak expression of a 1.8-kb mRNA in a few melanoma cell lines. Northern blot and RT-PCR analyses with DNA probes and oligonucleotide primers that correspond to distinct regions of full-length MLSN1 mRNA indicated that the short transcripts contained sequences corresponding primarily to either 5'- or 3'-end of the 5.4-kb mRNA. HMBA appears to up-regulate MLSN1 transcripts derived mainly from the 5'-end. Modulators of cAMP and protein kinase C pathways had no significant effect on MLSN1 expression. Our data show that multiple MLSN1 transcripts, both constitutively expressed and inducible, are present in cultured pigmented melanoma cells, and suggest that MLSN1 expression can be regulated at the level of both transcription and mRNA processing.