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Enhanced expression of bcl-2 inhibits apoptosis in cultured human keratinocytes
1Department of Dermatology, Kurume University School of Medicine, Japan.
Abstract:
The molecular mechanisms regulating epidermal differentiation and apoptosis have not been elucidated. Bcl-2, one of the candidate genes for suppressing apoptosis, was originally cloned from the breakpoint of at (14;18) translocation present in many human B cell lymphomas. In this study, the influence of bcl-2 on apoptosis was observed in transfected keratinocytes. After transfection of pEF-BOS vector with/without bcl-2, the expression of coded protein and the viability under starved conditions were examined. The bcl-2-transfected keratinocytes had cytoplasmic positive staining with anti-bcl-2 monoclonal antibodies, however the vector only transfected cells were devoid of the reaction products. The viability of transfected keratinocytes under starved conditions, with a lack of epidermal growth factor and bovine pituitary extract, was maintained in bcl-2 transfected cells; while the vector only transfected cells showed apoptotic cell death. The present result indicates that bcl-2 suppresses apoptotic cell death under starved conditions due to a lack of epidermal growth factor and bovine pituitary extract.
Insights
The gene BCL2 (B-cell lymphoma 2) prevents apoptosis (programmed cell death) in skin cells. BCL2-transfected keratinocytes survived starvation, unlike control cells, indicating BCL2
Area of Science:
- Molecular Biology
- Cell Biology
- Dermatology
Background:
- Molecular mechanisms of epidermal differentiation and apoptosis remain unclear.
- BCL2 (B-cell lymphoma 2) is a candidate gene for apoptosis suppression, identified in human B cell lymphomas.
Purpose of the Study:
- To investigate the influence of BCL2 on apoptosis in keratinocytes.
- To examine BCL2 protein expression and cell viability under nutrient-deprived conditions.
Main Methods:
- Transfection of keratinocytes with a vector containing or lacking the BCL2 gene.
- Immunohistochemical analysis using anti-BCL2 monoclonal antibodies to detect protein expression.
- Assessment of cell viability under starved conditions (lack of epidermal growth factor and bovine pituitary extract).
Main Results:
- BCL2 protein was detected in the cytoplasm of transfected keratinocytes.
- Vector-only transfected cells showed no BCL2 expression.
- BCL2-transfected keratinocytes maintained viability during starvation, while vector-only cells underwent apoptosis.
Conclusions:
- BCL2 gene expression suppresses apoptotic cell death in keratinocytes.
- This effect is evident under conditions of nutrient deprivation, specifically lacking epidermal growth factor and bovine pituitary extract.