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Identification and temporal expression pattern of genes modulated during irreversible growth arrest and terminal
Abstract:
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.
Insights
Treatment with fibroblast interferon (IFN-beta) and mezerein (MEZ) induces terminal differentiation in melanoma cells. This study identified 37 cDNAs, including novel genes, associated with this cancer cell growth control.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Cancer cell differentiation is frequently abnormal.
- Recombinant human fibroblast interferon (IFN-beta) and mezerein (MEZ) induce terminal differentiation and suppress proliferation in human melanoma cells.
- This differentiation is linked to differential gene expression regulating cancer cell growth and differentiation.
Purpose of the Study:
- To identify genes involved in melanoma cell differentiation and growth control.
- To understand the molecular mechanisms underlying IFN-beta and MEZ-induced melanoma cell differentiation.
Main Methods:
- Utilized a differentiation induction subtraction hybridization (DISH) scheme.
- Constructed a temporally spaced subtracted differentiation inducer treated (TSS) cDNA library.
- Employed high-throughput microchip cDNA array screening (Synteni) and Northern blotting for gene expression analysis.
Main Results:
- Screened approximately 10% of the DISH library (1000 clones).
- Identified and cloned 26 known and 11 novel cDNAs.
- These cDNAs are potentially relevant to growth control and terminal differentiation in human melanoma cells.
Conclusions:
- The study successfully identified key genes, including novel ones, associated with melanoma cell differentiation.
- This research provides a foundation for understanding the genetic regulation of melanoma cell growth and differentiation.