Proteomic analysis of the resistance to aplidin in human cancer cells.
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.
González-Santiago
Laura
L
Instituto de Investigaciones Biomédicas Alberto Sols, Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, Arturo Duperier, 4, E-28029 Madrid, Spain.
Alfonso
Patricia
P
Suárez
Yajaira
Y
Núñez
Antonio
A
García-Fernández
Luis F
LF
Alvarez
Enrique
E
Muñoz
Alberto
A
Casal
J Ignacio
JI
D000970:Antineoplastic Agents / Q000494:pharmacology*; D051017:Apoptosis Regulatory Proteins / Q000032:analysis*; D015153:Blotting, Western; D002462:Cell Membrane / Q000737:chemistry; D003600:Cytosol / Q000737:chemistry; D047630:Depsipeptides / Q000494:pharmacology*; D019008:Drug Resistance, Neoplasm; D015150:Echocardiography, Doppler; D006367:HeLa Cells; D006801:Humans; D009363:Neoplasm Proteins / Q000032:analysis*; D009369:Neoplasms / Q000737:chemistry*; D010456:Peptides, Cyclic; D040901:Proteomics
D000970:Antineoplastic Agents; D051017:Apoptosis Regulatory Proteins; D047630:Depsipeptides; D009363:Neoplasm Proteins; D010456:Peptides, Cyclic; C098980:plitidepsin