Identification and temporal expression pattern of genes modulated during irreversible growth arrest and terminal differentiation in human melanoma cells.
Abnormalities in differentiation are common occurrences in human cancers. Treatment of human melanoma cells with the combination of recombinant human fibroblast interferon (IFN-beta) and the antileukemic compound mezerein (MEZ) results in a loss of tumorigenic potential that correlates with an irreversible suppression in proliferative ability and induction of terminal differentiation. It is hypothesized that this is associated with the differential expression of genes that may directly regulate cancer cell growth and differentiation. To define the relevant gene expression changes that correlate with and potentially control these important cellular processes a differentiation induction subtraction hybridization (DISH) scheme is being used. A temporally spaced subtracted differentiation inducer treated (TSS) cDNA library was constructed and differentially expressed DISH clones were isolated and evaluated using a high throughput microchip cDNA (Synteni) array screening approach. Verification of differential gene expression for specific cDNAs was confirmed by Northern blotting. The temporal kinetics of regulation and the expression pattern of DISH genes were also evaluated by microchip cDNA array screening. Using this approach with 1000 DISH cDNA clones (approximately 10% of the DISH library) has resulted in the identification and cloning of both 26 known and 11 novel cDNAs of potential relevance to growth control and terminal differentiation in human melanoma cells.
Huang
F
F
GenQuest Incorporated, New York, NY 10032, USA.
Adelman
J
J
Jiang
H
H
Goldstein
N I
NI
Fisher
P B
PB
D002454:Cell Differentiation / Q000235:genetics*; D002455:Cell Division / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics* / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D015723:Gene Library; D006801:Humans; D008545:Melanoma / Q000235:genetics* / Q000473:pathology; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D004273:DNA, Neoplasm