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Analysis of the protein expression changes during taxol-induced apoptosis under translation inhibition conditions
David Piñeiro1, Víctor M González, Matilde Salinas
1Servicio Bioquímica-Investigación, Hospital Universitario Ramón y Cajal (Irycis), Ctra. Colmenar km 9,100, 28034, Madrid, Spain.
Abstract:
Taxol is currently used in chemotherapeutic treatments of different types of cancers. In this article, we demonstrate that taxol induces apoptosis and translation down-regulation in human embryonic kidney (HEK293T) cells. Antibody arrays are a promising new tool for the analysis of protein levels changes in cells responding to different stimuli. Using this approach, we have identified changes in the expression of 38 proteins (20 down-regulated and 18 up-regulated), implicated in several cellular processes mainly in apoptosis, cell cycle and signal transduction pathways, and also cytoskeleton proteins. Among them, we have confirmed a considerable decrease in the expression of p14(ARF) and a significant increase in the levels of dystrophin and c-Myc. It is known that c-Myc mRNA has an internal ribosome entry segment (IRES) element in its 5'UTR that could regulate its expression under global protein synthesis inhibition conditions. We demonstrate that after taxol treatment, the c-Myc IRES activity is maintained meanwhile cap-dependent activity is inhibited. In addition, an increase in c-Myc mRNA was also observed after taxol treatment. We conclude that taxol-induced c-Myc expression is regulated at both transcriptional and translational levels, the last of them by a mechanism mediated by IRES.
Insights
Taxol triggers apoptosis and reduces protein synthesis in kidney cells. This cancer drug alters 38 protein levels, including upregulating c-Myc via internal ribosome entry segment (IRES) activity.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- Taxol is a key chemotherapy agent for various cancers.
- Understanding Taxol's cellular mechanisms is crucial for improving cancer treatment.
- Protein expression analysis provides insights into drug responses.
Purpose of the Study:
- To investigate Taxol's effects on apoptosis and protein synthesis in human embryonic kidney (HEK293T) cells.
- To identify specific proteins and pathways affected by Taxol treatment.
- To elucidate the regulatory mechanism of c-Myc expression under Taxol-induced stress.
Main Methods:
- Utilized antibody arrays for high-throughput protein expression profiling.
- Analyzed changes in protein levels, focusing on apoptosis, cell cycle, and signal transduction pathways.
- Investigated c-Myc mRNA and internal ribosome entry segment (IRES) activity following Taxol treatment.
Main Results:
- Taxol induced apoptosis and global translation down-regulation in HEK293T cells.
- Identified 38 differentially expressed proteins, including decreased p14(ARF) and increased dystrophin and c-Myc.
- Confirmed Taxol treatment maintains c-Myc IRES activity while inhibiting cap-dependent translation, alongside increased c-Myc mRNA levels.
Conclusions:
- Taxol's effects extend to apoptosis induction and translational control.
- c-Myc expression is significantly modulated by Taxol at both transcriptional and translational levels.
- Internal ribosome entry segment (IRES)-mediated translation plays a key role in Taxol-induced c-Myc upregulation.
