Staurosporine induces apoptosis in NG108-15 cells
Bai-Fang Zhang1, Fang-Fang Peng, Jiang-Zhou Zhang
1Department of Biochemistry, School of Medicine, Wuhan University, Wuhan 430071, China.
Aim:
To observe if staurosporine induced apoptosis in NG108-15 cells and its effect on protein expression level of several genes related to apoptosis.
Methods:
Phase-contrast microscopy, fluorescence microscopy, and transmission electron microscopy were used to observe staurosporine-induced morphological changes. Agarose gel electrophoresis was used to detect DNA fragmentation. Western blots were used to measure protein expression level of several genes related to apoptosis.
Results:
Cells treated with staurosporine 0.1 micromol/L showed typical morphological changes of apoptosis. A "ladder" pattern representing fragmentation of DNA into oligonucleosome length fragments was observed after 6 h of staurosporine treatment and sustained until 24 h. The Bax expression increased significantly at 6 h after exposure to staurosporine, peaked at 12 h compared with vehicle cultures, and decreased at 24 h. The Bcl-2 expression increased and reached the highest level at 3 h. It was then decreased gradually but still higher than normal expression level. There was an obvious caspase-3 cleavage at 6 h after exposing the cells to staurosporine. Treatment with staurosporine for 12 h markedly decreased the expression of p53 protein. Cdk5 protein expression did not have obvious changes after the cells were exposed to staurosporine.
Conclusion:
Staurosporine induced apoptotic death in NG108-15 cells. Cells might die via a pathway that is dependent on Bax expression but independent of p53, and caspase-3 cleavage was involved.
Insights
Staurosporine triggers apoptosis in NG108-15 cells, evidenced by DNA fragmentation and altered protein levels of Bax, Bcl-2, and caspase-3. This cell death pathway appears independent of p53 expression.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Apoptosis is a crucial process for cellular homeostasis.
- Understanding the molecular mechanisms of apoptosis is vital for disease research.
- NG108-15 cells are a commonly used neuronal cell line model.
Purpose of the Study:
- To investigate staurosporine-induced apoptosis in NG108-15 cells.
- To analyze the impact of staurosporine on the protein expression of apoptosis-related genes.
Main Methods:
- Morphological analysis using phase-contrast, fluorescence, and transmission electron microscopy.
- Detection of DNA fragmentation via agarose gel electrophoresis.
- Quantification of protein expression using Western blot analysis.
Main Results:
- Staurosporine induced characteristic apoptotic morphological changes and DNA fragmentation.
- Bax expression increased, Bcl-2 expression showed initial increase then gradual decrease, and caspase-3 cleavage was observed.
- p53 protein expression decreased, while Cdk5 expression remained unchanged.
Conclusions:
- Staurosporine effectively induces apoptosis in NG108-15 cells.
- The apoptotic pathway involves Bax modulation and caspase-3 activation, potentially independent of p53.
- This study provides insights into the molecular regulation of staurosporine-induced apoptosis.
