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Identification of differentially expressed genes in organ-confined prostate cancer by gene expression array
1Department of Medicine, University of Sydney, Sydney, NSW, Australia.
Background:
To understand the molecular mechanisms underlying prostate cancer, we have utilized the gene expression array to search for genes whose expression is altered in this disease.
Methods:
RNA quality from manual microdissected tissue was compared with that from microselected tissue by electrophoresis. For array analysis, malignant and normal prostate epithelium was enriched using microselection technique from prostate cancer and the peripheral zone of a normal prostate. Identical array membrane was hybridized to labeled cancer and normal cDNA, respectively. The differentially expressed gene was further evaluated by RT-PCR.
Results:
Microdissection, but not microselection, causes visible degradation to RNA. Of the 588 genes on the membrane, 87 genes yielded significant signals. Based on a three fold difference relative to normal prostate tissue, 1 gene was overexpressed and 12 genes underexpressed in prostate cancer. Of them, five showed statistically significant reduction in mRNA levels in six prostate cancer specimens compared with seven normal prostate specimens. These five genes are glutathione S-transferase M1 (GSTM1), monocyte chemotactic protein-1 (MCP-1), tumor necrosis factor-alpha receptor-1 (TNFR-1), transforming growth factor beta3 (TGF-beta3), and inhibitor of DNA binding-1 (ID-1).
Conclusions:
GST-based metabolism, cytokine MCP-1 and TNFR-1, and TGF-beta3 signaling pathways, and some helix-loop-helix nuclear proteins could be potentially important in organ-confined prostate cancer and deserve further investigation.
Insights
Researchers identified 12 underexpressed genes in prostate cancer, including glutathione S-transferase M1 (GSTM1) and monocyte chemotactic protein-1 (MCP-1), potentially key to disease mechanisms.
Area of Science:
- Molecular Biology
- Oncology
- Genetics
Background:
- Prostate cancer molecular mechanisms remain incompletely understood.
- Gene expression profiling is a valuable tool for identifying disease-related alterations.
Purpose of the Study:
- To identify genes with altered expression in prostate cancer using gene expression arrays.
- To elucidate molecular mechanisms underlying prostate cancer development.
Main Methods:
- Comparison of RNA quality from manual microdissection versus microselection.
- Enrichment of prostate cancer and normal prostate epithelium using microselection.
- Hybridization of cDNA from cancer and normal tissues to identical gene arrays.
- Validation of differentially expressed genes using Reverse Transcription Polymerase Chain Reaction (RT-PCR).
Main Results:
- Microdissection, unlike microselection, degrades RNA.
- Out of 588 genes analyzed, 87 showed significant signals.
- One gene was overexpressed, and 12 were underexpressed in prostate cancer.
- Five genes, including glutathione S-transferase M1 (GSTM1), monocyte chemotactic protein-1 (MCP-1), tumor necrosis factor-alpha receptor-1 (TNFR-1), transforming growth factor beta3 (TGF-beta3), and inhibitor of DNA binding-1 (ID-1), showed statistically significant mRNA reduction.
Conclusions:
- Glutathione S-transferase (GST)-based metabolism, cytokine MCP-1, TNFR-1, and TGF-beta3 signaling pathways are potentially significant in organ-confined prostate cancer.
- Certain helix-loop-helix nuclear proteins may also play a role in prostate cancer.
- These findings warrant further investigation into the identified pathways and genes.