Study of the development of chemoresistance in melanoma cell lines using proteome analysis

Abstract

Malignant melanomas have poor prognosis since treatment with anti-neoplastic agents is mostly ineffective. The biological mechanisms of this strong intrinsic therapy resistance are unknown. In order to identify new molecular factors potentially associated with the drug-resistant phenotype of malignant melanoma, a panel of human melanoma cell variants exhibiting low and high levels of resistance to four commonly used anticancer drugs in melanoma treatment, i.e., vindesine, etoposide, cisplatin, and fotemustine, was characterized using proteomic tools (two-dimensional electrophoresis for protein fractionation and matrix assisted laser desorption/ionization-time of flight (MALDI-TOF)-mass spectrometry for protein identification). In the neutral and weak acidic milieu (pH 4.0-8.0) a total number of 14 proteins showed alterations in expression whereas 20 proteins were differentially expressed in the basic milieu (pH 8.0-11.0). Besides proteins with unknown physiologic function, several factors were identified that show chaperone activity. Moreover, proteins involved in drug detoxification, metabolism, and regulation of apoptotic pathways could be identified. The possible role of these proteins in the development of chemoresistance is discussed, although detailed functional tests with these proteins have still to be performed. Nevertheless, it is clear that this proteomic approach for studying chemoresistance phenomena is a prerequisite before further investigation can yield insight into the biology and development of drug resistance in malignant melanoma.

Microscale solution IEF combined with 2‐D DIGE substantially enhances analysis depth of complex proteomes such as mammalian cell and tissue extracts

“…3A) from a total of 1658 spots. The number of protein spots detected here on 2-D gels is similar to results from other proteomic studies of melanoma cell lines where about 1000-2000 spots were detected by 2-D gels [29][30][31][32].…”

Quantitative proteome analysis of cisplatin‐induced apoptotic Jurkat T cells by stable isotope labeling with amino acids in cell culture, SDS‐PAGE, and LC‐MALDI‐TOF/TOF MS

“…Furthermore, cisplatin has a number of side effects, such as neurotoxicity and nephrotoxicity, and presence or acquisition of resistance limits its use [9]. Thus, several proteome analyses with different cell lines were performed to identify proteins associated with resistance to cisplatin [10][11][12][13][14].…”

A systems biology analysis of metastatic melanoma using in‐depth three‐dimensional protein profiling

“…In addition, there may be some similarities in biological mechanisms implicated in melanoma progression and those that play a role in development of drug resistance in malignant melanoma. For example, proteins involved in chemoresistance of malignant melanoma cells, such as elongation factor 1-delta, translationally controlled tumor protein, 60 kDa heat shock protein, and nucleophosmin [23,24] were differentially expressed in metastatic melanoma.…”