Correlation of genetic instability and apoptosis in the presence of oncogenic Ki-Ras
Related Experiment Videos
1Cancer Research Center, Boston University School of Medicine, Massachusetts 02118, USA.
Abstract:
The product of the ras proto-oncogene has been implicated as an essential signal transducer, involved in a variety of biological or pathological activities, including apoptosis. The aim of this investigation was to further explore the mechanisms of apoptosis triggered by Ras. Stable expression of constitutively-activated (v)-Ki-Ras in Balb/c-3T3 mouse fibroblasts resulted in a loss of G1 arrest in response to treatments which induced cell cycle arrest in the parental Balb/c-3T3 cells, accompanied by decreased expression of the p53 tumor suppressor protein and the GADD45 gene, the product of which is involved in DNA repair, and deregulated expression of the MDM-2 gene, the product of which can regulate p53 expression. Ki-Ras expression also increased the frequency of PALA-selectable CAD gene amplification, and paradoxically the susceptibility to PALA-induced apoptosis. After persistent serum-starvation, cells expressing the activated ras gene lost clonogenic potential, indicating impaired capability for genetic repair in the cells. Taken together, these data suggest that activated Ki-ras may confer genetic instabilty upon cells, possibly through interference with tumor suppressors, such as p53. While this instability may facilitate adaptation to environmental stresses, this instability in the genome also renders cells containing activated ras genes intrinsically more susceptible to programmed cell death, possibly by accumulation of undesirable or lethal genetic events during the process of tumor development.
Insights
Activated Ras signaling in mouse fibroblasts disrupts cell cycle control and DNA repair, paradoxically increasing susceptibility to apoptosis. This suggests Ras may drive genetic instability, impacting tumor development and cell death.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Ras proto-oncogenes are crucial signal transducers in biological and pathological processes.
- Ras signaling is implicated in diverse cellular activities, including programmed cell death (apoptosis).
Purpose of the Study:
- To investigate the mechanisms by which Ras triggers apoptosis.
- To explore the role of activated Ki-Ras in cellular responses and genetic stability.
Main Methods:
- Stable expression of activated (v)-Ki-Ras in Balb/c-3T3 mouse fibroblasts.
- Analysis of cell cycle arrest, gene expression (p53, GADD45, MDM-2), gene amplification (CAD), and apoptosis.
- Assessment of clonogenic potential after serum starvation.
Main Results:
- Activated Ki-Ras expression led to loss of G1 arrest and decreased p53 and GADD45 expression.
- Deregulation of MDM-2 and increased CAD gene amplification were observed.
- Cells with activated Ki-Ras showed paradoxical susceptibility to apoptosis and impaired genetic repair, losing clonogenic potential.
Conclusions:
- Activated Ki-Ras may induce genetic instability by interfering with tumor suppressors like p53.
- This instability can aid adaptation but increases susceptibility to apoptosis due to accumulated genetic events.
- Ras-driven genetic instability is linked to both tumor development and programmed cell death.
Related Concept Videos
The Ras Gene
Ras is a superfamily...
The Intrinsic Apoptotic Pathway
Cancer-Critical Genes I: Proto-oncogenes
When the function of certain critical genes, especially those involved in cell cycle regulation and cell growth signaling cascades, gets disrupted, it upsets the cell cycle progression. Such cells with unchecked cell cycles start proliferating uncontrollably and eventually develop into tumors.
Such genes that act...
Small GTPases - Ras and Rho
Three regulatory proteins control their activity:
Abnormal Proliferation
The Retinoblastoma Gene
The first-ever tumor suppressor gene called Rb was identified in retinoblastoma - a rare eye tumor in children. In inherited forms of the disease, a child inherits one defective copy of the Rb gene, which predisposes them to retinoblastoma. However,...