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Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays
Published on: September 19, 2018
Proteomic analysis of the resistance to aplidin in human cancer cells
Laura González-Santiago1, Patricia Alfonso, Yajaira Suárez
1Instituto de Investigaciones Biomédicas Alberto Sols, Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, Arturo Duperier, 4, E-28029 Madrid, Spain.
Abstract:
Aplidin (plitidepsin) is an antitumoral agent that induces apoptosis via Rac1-JNK activation. A proteomic approach using 2D-DIGE technology found 52 cytosolic and 39 membrane proteins differentially expressed in wild-type and Aplidin-resistant HeLa cells, of which 39 and 27 were identified by MALDI-TOF mass spectrometry and database interrogation. A number of proteins involved in apoptosis pathways were found to be deregulated. Alterations in Rab geranylgeranyltransferase, protein disulfide isomerase (PDI), cystathionine gamma-lyase, ezrin, and cyclophilin A (CypA) were confirmed by immunoblotting. Moreover, the role of PDI and CypA in Aplidin resistance was functionally confirmed by using the inhibitor bacitracin and overexpression, respectively. These deregulated proteins are candidates to mediate, at least partially, Aplidin action and might provide a route to the cells to escape the induction of apoptosis by this drug.
Insights
Aplidin (plitidepsin) induces cancer cell death by activating apoptosis. Researchers identified key proteins, like protein disulfide isomerase (PDI) and cyclophilin A (CypA), involved in resistance to this antitumoral agent.
Area of Science:
- Proteomics
- Cancer Biology
- Molecular Pharmacology
Background:
- Aplidin (plitidepsin) is an antitumoral agent targeting cancer cells.
- It induces apoptosis, a programmed cell death pathway, through Rac1-JNK signaling.
- Understanding mechanisms of resistance is crucial for optimizing cancer therapy.
Purpose of the Study:
- To investigate proteomic changes in HeLa cells resistant to Aplidin.
- To identify proteins involved in mediating Aplidin resistance and apoptosis evasion.
- To explore potential therapeutic targets for overcoming Aplidin resistance.
Main Methods:
- Proteomic analysis using 2D-DIGE technology.
- Protein identification via MALDI-TOF mass spectrometry and database interrogation.
- Validation of protein alterations using immunoblotting and functional assays (inhibitor treatment, overexpression).
Main Results:
- Differential expression of 52 cytosolic and 39 membrane proteins in resistant vs. wild-type cells.
- Identification of deregulated proteins, including Rab geranylgeranyltransferase, protein disulfide isomerase (PDI), cystathionine gamma-lyase, ezrin, and cyclophilin A (CypA).
- Functional confirmation of PDI and CypA roles in Aplidin resistance.
Conclusions:
- Deregulated proteins, particularly PDI and CypA, are implicated in mediating Aplidin's antitumoral effects.
- These proteins may provide a mechanism for cancer cells to escape Aplidin-induced apoptosis.
- Targeting these identified proteins could offer strategies to overcome Aplidin resistance.
