Subtraction hybridization identifies a novel melanoma differentiation associated gene, mda-7, modulated during human melanoma differentiation, growth and progression.

摘要: Cultured human melanoma cells lose proliferative capacity and terminally differentiate after treatment with the combination of recombinant fibroblast interferon (IFN-beta) mezerein (MEZ). Subtraction hybridization cDNA libraries prepared from actively proliferating H0-1 produced treated IFN-beta + MEZ identifies a novel differentiation-associated (mda) cDNA, mda-7, that displays elevated expression in differentiation inducer-treated cells. mda-7 encodes protein 206 amino acids predicted size 23.8 kDa. The level mRNA is normal melanocytes versus primary metastatic melanomas. In Matrigel-assisted progression model, decreases early vertical growth phase selected for autonomous or enhanced tumor formation nude mice. Treatment melanomas MEZ, to lesser extent results suppression induced expression. Immunoprecipitation analyses using peptide-derived rabbit polyclonal antibodies detect increases protein, higher molecular weight approximately 90 100 kDa, highly conserved gene an homologous sequence genome yeast. Transfection constructs into C8161 reduces inhibits colony formation. These confirm has antiproliferative properties this context may contribute terminal cell differentiation. also function as negative regulator progression.