DFNA5 (ICERE-1) contributes to acquired etoposide resistance in melanoma cells.
Resistance to drug treatment is a common observation in malignant melanoma. In order to analyze alterations in mRNA expression profiles associated with drug resistance in melanoma cells we previously established a panel of various drug-resistant cell variants derived from the human melanoma line MeWo and compared the mRNA expression profiles by a differential display technique. By that approach it could be demonstrated that the expression level of a mRNA encoded by a gene found to be mutated in non-syndromic hearing impairment, DFNA5 (ICERE-1), was distinctly decreased in the 33-fold etoposide-resistant melanoma cell line MeWo ETO 1. To evaluate the hypothesis that a decrease in DFNA5 mRNA expression level contributes to the acquired etoposide resistance phenotype exhibited by MeWo ETO 1 cells, this drug-resistant line was stably transfected with the DFNA5-encoding cDNA. Transfected clones showed a 30-35% reduced etoposide susceptibility by comparing the IC(25), IC(50) and IC(75) values of these clones with those displayed by the non-transfected, etoposide-resistant melanoma cell line MeWo ETO 1 and controls. Furthermore, etoposide exposure of stable DFNA5 transfectants resulted in an increase of caspase-3-mediated apoptotic events in DFNA5-transfected clones in comparison to MeWo ETO 1 cells and controls. The data therefore demonstrate that a decrease in DNFA5 mRNA expression level is associated with an increased etoposide resistance in melanoma cells due to an elevated cellular susceptibility to trigger a caspase-3-depending signal pathway leading to programmed cell death.
Lage
H
H
Institute of Pathology, Charité, Campus Mitte, Humboldt University Berlin, Germany. hermann.lage@charite.de
Helmbach
H
H
Grottke
C
C
Dietel
M
M
Schadendorf
D
D
D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D017209:Apoptosis; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D019008:Drug Resistance, Neoplasm; D004789:Enzyme Activation; D005047:Etoposide / Q000494:pharmacology*; D015870:Gene Expression; D006801:Humans; D008545:Melanoma / Q000188:drug therapy*; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D014162:Transfection; D014407:Tumor Cells, Cultured; D000094524:Gasdermins
D000972:Antineoplastic Agents, Phytogenic; D002352:Carrier Proteins; D053148:Caspase 3; D020169:Caspases; D005047:Etoposide; D012333:RNA, Messenger; D011960:Receptors, Estrogen; C111432:GSDME protein, human; D000094524:Gasdermins; C505407:CASP3 protein, human