Characterization of human melanoma cell lines and melanocytes by proteome analysis

Abstract

We have analyzed the proteomes of two human melanoma cell lines (A375 and 526), and of the human melanocytes, (FOM 78), by two-dimensional electrophoresis (2D-PAGE) and liquid chromatography - tandem mass spectrometry (LC-MS/MS). Our comparative proteomic analysis revealed that six proteins were over-expressed in both melanoma cell lines as compared to melanocytes: galectin-1, inosine-5'-monophosphate dehydrogenase 2, serine/threonine-protein phosphatase 2A 65 kDa regulatory subunit A alpha isoform, protein DJ-1, cyclophilin A and cofilin-1. We show, for the first time, that only specific isoforms of these molecules are over-expressed in melanoma. Different protein profiles were also found between each individual melanoma cell line and the melanocytes. s-Methyl-5-thioadenosine phosphorylase, ubiquitin and ribosomal protein S27 a precursor, the basic form of protein DJ-1, annexin a1, proliferation associated protein 2g4, isoform alfa-enolase of alfa-enolase, protein disulfide-isomerase precursor, and elongation factor 2 were more strongly expressed in A375 cells compared to melanocytes. In 526 cells, 60s acidic ribosomal protein p1 and calreticulin precursor were more highly expressed than in melanocytes. These molecular differences may help in better understanding melanoma development and its different responsiveness to therapies. The identified proteins could be exploited as biomarkers or therapeutic targets for melanoma.

Rapid and Deep Human Proteome Analysis by Single-dimension Shotgun Proteomics

“…A375 has been widely used in cytokine research (mostly IL1 response studies), as well as melanoma cancer research (26). There have been several transcriptomics and proteomics studies of the cell line (27,28); however, until now there has been no report published on the deep proteome profile of this cell line.…”

Double amplifications of CDK4 and MDM2 in a gastric inflammatory myofibroblastic tumor mimicking cancer with local invasion of the spleen and diaphragm

“…CDK4 and the cyclin D1 complex are inhibited by INK4 proteins, which include p16 ink4a , p17 ink4b , p18 ink4c , and p19 ink4d. 23,24 Under strong oncogenic stress, p16 ink4a overexpression ultimately regulates RB protein to suppress growth and cell cycle progression, which promotes oncogene-induced senescenc. 23,24 In certain pre-malignant lesions, high levels of p16 ink4a are observed and are believed to contribute to arresting the lesions' progression.…”

PHF19 and Akt control the switch between proliferative and invasive states in melanoma

“…27), and recent results, obtained by proteomic analysis, showed an increase of PP2A expression (one of the known phosphatases of Akt) and of DJ1 (which regulates PTEN activity) and suggest a modification of regulation of Akt activity in melanoma cells. 28 We hypothesized that Akt might act upstream PHF19 to regulate both PHF19 expression and the pluripotency genes (see Fig. 8 for a schematic view of these interactions).…”