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Detection of differentially expressed genes in prostate cancer by combining suppression subtractive hybridization and
1Laboratory of Cancer Genetics, Institute of Medical Technology, University of Tampere and Tampere University Hospital, Tampere, Finland.
Abstract:
The molecular mechanisms underlying the development and progression of prostate cancer have remained poorly understood. The identification of differentially expressed genes has been used as a tool to recognize genes that are involved in disease processes. In this study we combined suppression subtractive hybridization (SSH) and cDNA array hybridization to identify genes whose expression is decreased in prostate cancer. cDNA from benign prostatic hyperplasia (BPH) was subtracted with cDNA from the prostate cancer cell line PC-3 and 386 of the subtracted clones were arrayed onto a nylon filter membrane. The differential gene expression was then verified by hybridizing the filter with radioactively labelled first-strand cDNA preparations from BPH, PC-3, four other cancer cell lines, and a normal prostate epithelial cell line (PrEC). In order to validate SSH and cDNA array hybridization, the enrichment of clones in the subtraction, as well as the sensitivity and linearity of array hybridization, was first evaluated. The array hydridization results were confirmed by northern analysis and selected clones were sequenced. Altogether, several known genes, such as prostate-specific antigen (PSA), human glandular kallikrein 2 (hK2), phosphatidic acid phosphatase type 2a (PAP2a), alpha-tropomyosin, and insulin-like growth factor binding protein 7 (IGFBP-7), as well as an anonymous transcript (EST), were found to be expressed less in PC-3 than in BPH. Further studies on the significance of these genes in the development of prostate cancer are now warranted.
Insights
Researchers identified genes with decreased expression in prostate cancer using suppression subtractive hybridization (SSH) and cDNA array analysis. This study helps understand prostate cancer development by pinpointing key genes like PSA and hK2.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Prostate cancer development and progression mechanisms are not fully understood.
- Identifying differentially expressed genes aids in recognizing disease-involved genes.
Purpose of the Study:
- To identify genes with decreased expression in prostate cancer using SSH and cDNA array hybridization.
- To understand the molecular mechanisms of prostate cancer.
Main Methods:
- Suppression subtractive hybridization (SSH) combined with cDNA array hybridization.
- Subtraction of benign prostatic hyperplasia (BPH) cDNA from prostate cancer cell line PC-3 cDNA.
- Verification of differential gene expression using northern analysis and sequencing.
Main Results:
- Identified several known genes (PSA, hK2, PAP2a, alpha-tropomyosin, IGFBP-7) and an EST with decreased expression in PC-3 compared to BPH.
- Validated SSH and cDNA array hybridization methods for accuracy and sensitivity.
Conclusions:
- Several genes show reduced expression in prostate cancer cells, suggesting their potential role in disease development.
- Further research is warranted to explore the significance of these identified genes in prostate cancer.