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Abnormal prostate development in C3(1)-bcl-2 transgenic mice
1Department of Urology, College of Physicians and Surgeons, Columbia University, New York, New York 10032, USA.
Background:
Recent hypotheses to explain the etiology of abnormal growth associated with prostate disease have invoked perturbations in the rate of apoptosis as an important contributor to the onset and progression of these diseases. For this reason, the apoptosis suppressing oncoprotein bcl-2 has come under scrutiny with regards to its role in prostate diseases. In order to evaluate the role of bcl-2 in human prostate disease and to develop an animal model to test anti-bcl-2 therapies, we generated transgenic mice in which bcl-2 expression is targeted to the mouse prostate gland.
Methods:
Mouse embryos were microinjected with recombinant DNA constructed by fusing a modified rat C3(1) promotor element to cDNA encoding human bcl-2. Presence of the C3(1)-bcl-2 transgene in progeny was identified by Southern blot and polymerase chain reaction (PCR) analysis. RNase protection assays were used to analyze RNA from 15 organs of these mice. Western blot assays and immunohistochemical staining were used to confirm the tissue-specific protein expression of human bcl-2 and its cellular localization.
Results:
Three lines of C3(1)-bcl-2 transgenic mice were established. Founder mice carried 2-20 copies of the transgene. Expression of human bcl-2 from the transgene was limited to the prostate gland and testis of males as well as the uterus of females. In the prostate gland, human bcl-2 protein was found only in prostatic epithelial cells. Microscopic analysis of prostate glands from individual males (three lines) showed that these glands were often abnormal, with increased accumulation of cells in the prostatic stroma as well as the epithelium.
Conclusions:
These transgenic mice appear to provide a novel animal model for studying neoplastic development of the prostate, with particular emphasis on the bcl-2 protein and the role of apoptosis regulation in such development.
Insights
Researchers created transgenic mice to study prostate disease. These mice overexpress bcl-2 in the prostate, showing abnormal growth and providing a new model for investigating apoptosis regulation in neoplastic development.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Abnormal prostate growth is linked to apoptosis dysregulation.
- The bcl-2 oncoprotein, which suppresses apoptosis, is implicated in prostate diseases.
- Understanding bcl-2's role is crucial for developing targeted therapies.
Purpose of the Study:
- To investigate the role of bcl-2 in human prostate disease.
- To develop a transgenic mouse model for studying prostate neoplastic development.
- To test potential anti-bcl-2 therapies.
Main Methods:
- Generated transgenic mice by microinjecting a C3(1)-bcl-2 construct into embryos.
- Confirmed transgene presence using Southern blot and PCR.
- Analyzed bcl-2 expression at RNA and protein levels via RNase protection, Western blot, and immunohistochemistry.
Main Results:
- Established three lines of C3(1)-bcl-2 transgenic mice.
- Human bcl-2 expression was specific to the prostate gland, testis, and uterus.
- Prostate glands exhibited abnormal growth with increased cell accumulation in stroma and epithelium.
Conclusions:
- The developed transgenic mice serve as a novel model for prostate neoplastic development.
- This model facilitates the study of bcl-2's role in apoptosis regulation.
- It offers a platform for evaluating anti-bcl-2 therapeutic strategies.