Related Experiment Videos
Apoptosis induced by protein kinase C inhibition in a neuroblastoma cell line
M M Behrens1, J L Martínez, C Moratilla
1Institute of Biomedical Investigations, Cosejo Superior de Investigaciones Científicas, CSIC, Department of Biochemistry, Universidad Autónoma de Madrid, Spain.
Abstract:
The protein kinase C inhibitor bisindolylmaleimide GF109203X has a dual effect on the behavior of the neuroblastoma cell line Neuro-2A; when the inhibitor is added in conditions that induce differentiation (absence of serum), neurite outgrowth is potentiated in a dose-dependent manner. However, if the inhibitor is added in growth-promoting conditions (presence of serum), programmed cell death (apoptosis) is induced, as assessed by internucleosomal DNA cleavage and specific immunoassays. This effect is also seen with other specific protein kinase C inhibitors. Bcl2 gene overexpression protects Neuro-2A cells from apoptosis, as has been found in other systems. We also show that calpain I, a neutral Ca(2+)-activated proteinase, participates in this apoptotic pathway. Our results point to a key role of protein kinase C in the regulation of growth and differentiation in Neuro-2A cells.
Insights
Protein kinase C inhibition differentially affects Neuro-2A cells. It promotes neurite outgrowth during differentiation but induces apoptosis in growth conditions, involving calpain I.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Protein kinase C (PKC) plays a role in cellular processes.
- Neuroblastoma cell line Neuro-2A is a model for neuronal development and cancer.
Purpose of the Study:
- To investigate the dual role of PKC inhibition in Neuro-2A cell behavior.
- To elucidate the molecular mechanisms underlying PKC inhibitor effects on differentiation and apoptosis.
Main Methods:
- Treatment of Neuro-2A cells with bisindolylmaleimide GF109203X under varying serum conditions.
- Assessment of neurite outgrowth for differentiation.
- Analysis of internucleosomal DNA cleavage and immunoassays for apoptosis.
- Evaluation of Bcl2 gene overexpression and calpain I activity.
Main Results:
- Bisindolylmaleimide GF109203X potentiated neurite outgrowth in serum-free conditions (differentiation).
- The inhibitor induced apoptosis in serum-containing conditions (growth).
- Bcl2 overexpression conferred protection against apoptosis.
- Calpain I was identified as a participant in the apoptotic pathway.
Conclusions:
- Protein kinase C inhibition has opposing effects on Neuro-2A cell differentiation and apoptosis.
- PKC is a key regulator of growth and differentiation in Neuro-2A cells.
- Calpain I is involved in the PKC-mediated apoptotic pathway.