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Identification of mRNAs and proteins specifically associated with transformation in ocular malignant melanoma cells
Author(s) -
COSTAGLIOLA C,
SEMERARO F,
ROMANO MR,
CAPUTO M,
DE ROSA MC,
TECCE MF
Publication year - 2012
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2012.s013.x
Subject(s) - melanoma , malignant transformation , transfection , biology , ocular melanoma , cancer research , cell culture , messenger rna , cell , gene , microbiology and biotechnology , genetics
Purpose Ocular malignant melanoma has several morphological specificity when compared to melanoma tissues from other body regions. To assess and evaluate specific molecular factors of ocular melanoma we decided to screen clinical specimens from patients with melanoma from different tissue and organs, including eye. Methods To identify a number of factors to be analyzed in clinical samples, we selected mRNAs specifically associated with the transformation state of cultured human melanoma cell line A375 This cell line was used as typical reference status of melanoma cells. These cells were then compared to similar cells in which Bag3 mRNA was silenced by transfection with specific siRNA (small interfering for Bag3). This mRNA encodes a protein which has a potent anti‐apoptotic effect and is generally expressed in melanoma cells, being part of the molecular mechanism of their transformation status. Results Microarray techniques took advantage to compare the whole pattern of cellular mRNAs in the two conditions of A375 cells: untreated neoplastic and Bag3 silenced cells. Several mRNA species resulted differentially expressed between the two conditions both as induced or as decreased when comparing the more transformed condition vs. Bag3 silenced cells. We then tested the expression of some of these mRNA and of their encoded proteins within the clinical samples of ocular and non‐ocular melanoma tissues. Conclusion Preliminary results indicated the expression of nephroblastoma overexpressed gene (NOV) as typical of melanoma transformation and of a small pattern of proteins as selectively modulated in ocular melanoma.