Regulation of rat DOC-2 gene during castration-induced rat ventral prostate degeneration and its growth inhibitory function in human prostatic carcinoma cells
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Updated: Oct 29, 2026
Murine Prostate Micro-dissection and Surgical Castration
Published on: May 11, 2016
1Department of Urology, University of Texas Southwestern Medical Center, Dallas 75235-9110, USA.
Abstract:
Androgen is a mitogen as well as a morphogen for prostatic epithelium. However, the detailed mechanisms of these distinct androgenic actions have not yet been delineated. Therefore, we employed differential display PCR to unveil any potential genes that may be involved in these processes. In this study, we report the isolation and characterization of two alternative splicing forms (p82 and p59) of C9 complementary DNA, the rat homolog of the human deletion of ovarian carcinoma 2 (DOC-2) gene and mouse p96 phosphoprotein, from rat ventral prostate (VP). We found that C9 was up-regulated in rat VP after castration, suggesting that C9 may be regulated by androgen receptor directly or indirectly during prostate degeneration. A similar regulatory pattern was also observed in both the seminal vesicle and dorsolateral prostate, but not in the coagulating gland or other androgen-independent organs. Immunohistochemical analysis of rat VP demonstrated that C9 is detected in the basal epithelia and surrounding stromal cells after prolonged castration. Ribonuclease protection assay and Western blot analysis revealed that p59 is the predominant C9 isoform in rat VP. To unveil the function of C9 in cell growth, we transfected p59 complementary DNA into the C4-2 cells, a derivative of the LNCaP prostatic carcinoma cell line. The p59 stable transfectants exhibited a slower growth rate and an increase in the cell fraction in the G1 phase under our experimental conditions. These data indicate that C9-p59 has growth inhibitory activity for prostatic epithelial cells. Taken together, our results suggest that C9 is up-regulated during prostate degeneration process and may play an active role in the proliferation and differentiation of prostatic epithelium.
Insights
Researchers identified C9, a gene regulated by androgens, which is upregulated in the prostate after castration. The C9-p59 form inhibits prostatic epithelial cell growth, suggesting a role in prostate degeneration and differentiation.
Area of Science:
- Molecular Biology
- Cell Biology
- Genetics
Background:
- Androgens are crucial for prostate development and function, acting as both mitogens and morphogens.
- The precise molecular mechanisms underlying androgenic actions in the prostate remain incompletely understood.
- Identifying novel genes involved in androgen-regulated prostate processes is essential.
Purpose of the Study:
- To identify genes involved in androgenic actions in the rat ventral prostate (VP).
- To characterize the function of the identified gene, C9, in prostatic epithelial cell growth.
Main Methods:
- Differential display PCR was used to identify potential genes.
- C9 complementary DNA (cDNA) was isolated and characterized.
- Immunohistochemistry, ribonuclease protection assay, and Western blot analysis were performed.
- C9-p59 cDNA was transfected into C4-2 cells to assess its effect on cell growth.
Main Results:
- Two alternative splicing forms (p82 and p59) of C9 cDNA were identified in rat VP.
- C9 expression was upregulated in rat VP post-castration, indicating androgen regulation.
- C9 is localized in basal epithelia and stromal cells after castration.
- The C9-p59 isoform was predominant in rat VP.
- Transfection with C9-p59 resulted in slower cell growth and G1 phase arrest in C4-2 cells.
Conclusions:
- C9 is an androgen-regulated gene in the rat prostate.
- The C9-p59 isoform exhibits growth inhibitory activity on prostatic epithelial cells.
- C9 may play a significant role in prostate degeneration, proliferation, and differentiation.
