ABSTRACT
The main role of growth arrest and DNA damage-inducible (GADD) genes is to block proliferation at G1 and G2 checkpoints in response to DNA damage. The goal of this study was to examine the expression of GADD genes in primary melanomas with respect to prognosis. GADD34 was found in 73% of the primary melanomas investigated. GADD45 and GADD153 were positive in 60% and 80% of primary melanomas, respectively. Cox regression demonstrated that only GADD153 had any independent prognostic impact. We therefore decided to establish a PCR assay for detection of GADD153 in paraffin-embedded tissue. GADD153 deletion was found in 3/26 melanomas. None of the 3 cases with GADD153 deletion showed any expression of GADD153. Sequencing analysis detected polymorphism T-C at amino acid position 10 in 6/23 melanomas. In 6 cases with GADD153 polymorphism, GADD153 expression was found in 2 melanomas with a maximum GADD153 index of 10%. We postulate that the GADD gene family plays an important role in melanoma progression.
Subject(s)
CCAAT-Enhancer-Binding Proteins/biosynthesis , Melanoma/diagnosis , Melanoma/metabolism , Proteins , Transcription Factors/biosynthesis , Adolescent , Adult , Aged , Aged, 80 and over , Antigens, Differentiation , Cell Cycle , Cell Cycle Proteins , Child , Disease-Free Survival , Genetic Techniques , Humans , Immunohistochemistry , Intracellular Signaling Peptides and Proteins , Melanoma/mortality , Middle Aged , Polymerase Chain Reaction , Polymorphism, Genetic , Prognosis , Proportional Hazards Models , Protein Biosynthesis , Protein Phosphatase 1 , Regression Analysis , Sequence Analysis, DNA , Time Factors , Transcription Factor CHOP , GADD45 ProteinsABSTRACT
The retinoblastoma gene is a cell cycle regulator preventing cells from entering into S-phase. An altered expression of the retinoblastoma gene has been reported in the majority of human malignancies. The main aim of this study was to investigate retinoblastoma gene expression in the full spectrum of melanoma progression from naevus to melanoma metastases by applying immunohistochemistry and RT-PCR. All naevi with and without dysplasia showed high expression of the retinoblastoma gene. In primary melanomas, Rb-positive cells were found in 82 out of 106. Loss of expression correlated with an increase in Clark level and shorter survival rates. An independent prognostic role of the retinoblastoma gene was confirmed by Cox multivariate analyses (p < 0.01). In melanoma metastases, retinoblastoma gene expression (at the RNA level) was found in 18 out of 26 melanoma lymphatic metastases, and in 2 out of 5 liver metastases. Our results indicate a downregulation of the retinoblastoma gene in the progression of melanocytic tumours.
Subject(s)
Genes, Retinoblastoma , Melanoma/metabolism , Retinoblastoma Protein/metabolism , Skin Neoplasms/metabolism , Adolescent , Adult , Aged , Aged, 80 and over , Child , Disease Progression , Down-Regulation , Female , Humans , Lymph Nodes/metabolism , Lymph Nodes/pathology , Lymphatic Metastasis/pathology , Male , Melanoma/mortality , Melanoma/secondary , Middle Aged , Neoplasm Staging , Nevus/genetics , Nevus/metabolism , Nevus/pathology , RNA, Neoplasm/analysis , Retinoblastoma Protein/genetics , Reverse Transcriptase Polymerase Chain Reaction , Skin Neoplasms/mortality , Skin Neoplasms/pathology , Survival RateABSTRACT
Thirty-one naevus cell naevi, 30 dysplastic naevi and 12 fibromatous naevi were stained for the presence of p53 and Growth Arrest DNA Damage genes. All naevus cell naevi and fibromatous naevi were highly positive for GADD genes and negative for p53. Dysplastic naevi had significantly lower GADD34 and GADD153 expression as well as higher p53 expression in relation to the other naevi groups. The absence or decrease of GADD genes expression in naevus may indicate a potential malignant transformation.
Subject(s)
CCAAT-Enhancer-Binding Proteins , DNA Damage/genetics , DNA, Neoplasm/genetics , Nevus/genetics , Skin Neoplasms/genetics , Adolescent , Adult , Aged , Aged, 80 and over , Antigens, Differentiation , Cell Cycle Proteins , Cell Division/genetics , Child , DNA, Neoplasm/metabolism , DNA-Binding Proteins/biosynthesis , DNA-Binding Proteins/genetics , Female , Genes, p53 , Humans , Intracellular Signaling Peptides and Proteins , Male , Middle Aged , Nevus/metabolism , Nevus/pathology , Protein Biosynthesis , Protein Phosphatase 1 , Proteins/genetics , Skin Neoplasms/metabolism , Skin Neoplasms/pathology , Transcription Factor CHOP , Transcription Factors/biosynthesis , Transcription Factors/genetics , GADD45 ProteinsABSTRACT
29 oral melanomas were stained immunohistochemically for the GADD34, GADD45 and GADD153. GADD34 was found in 5/29 melanomas and its expression did not exceed 21%, averaging 4.1% of melanoma cells. GADD45 was observed in 8/29 oral melanomas and cell positivity averaged 2.8%. GADD153 was found in 2/29 melanomas and the percentage of positive cells ranged between 0 and 31%, averaging 1.2%. Not one significant correlation between GADD genes in oral melanomas was found. Loss of GADD gene expression and lack of correlation between them show advanced disturbances in their cooperation, leading to a high genetic instability of oral melanoma cells.
Subject(s)
CCAAT-Enhancer-Binding Proteins , DNA Damage , Gene Expression Regulation, Neoplastic , Melanoma/genetics , Mouth Neoplasms/genetics , Adult , Aged , Aged, 80 and over , Antigens, Differentiation , Biomarkers, Tumor , Cell Cycle Proteins , Cell Nucleus , DNA-Binding Proteins/genetics , Female , Humans , Intracellular Signaling Peptides and Proteins , Male , Middle Aged , Protein Phosphatase 1 , Proteins/genetics , Transcription Factor CHOP , Transcription Factors/genetics , GADD45 ProteinsABSTRACT
The nm23 monoclonal antibody directed to specific antigen was applied for investigation of 29 benign naevi, 59 melanomas of different Clark level and locations, 4 lymph node and 28 organ metastases, using quantitative immunohistochemistry. The reduced expression of nm23 protein was associated with the tumor progression. A statistically significant difference (p = 0.0014) was found in nm23 protein expression between naevi and malignant melanomas. Additionally, a reduced protein level in metastases was found. A strong correlation was found between nm23 expression and survival of patients as well as the presence of metastasis (p = 0.0003).
Subject(s)
Biomarkers, Tumor/metabolism , Melanoma/metabolism , Monomeric GTP-Binding Proteins , Nucleoside-Diphosphate Kinase , Skin Neoplasms/metabolism , Transcription Factors/metabolism , Adolescent , Adult , Aged , Aged, 80 and over , Disease Progression , Female , Humans , Immunohistochemistry , Male , Melanoma/pathology , Middle Aged , NM23 Nucleoside Diphosphate Kinases , Nevus/metabolism , Nevus/pathology , Prognosis , Skin Neoplasms/pathologyABSTRACT
Twenty nine benign naevi and 59 melanomas of different Clark level and locations were stained immunohistochemically for the presence of p53 and 3 members of Growth Arrest. DNA Damage inducible family: GADD 34, GADD 45 and GADD 153. All naevi were p53 negative and highly GADD positive. Differences in the expression of all 3 GADD proteins between naevi and melanomas were highly significant (p = 0.0001). The expression of p53 increased and the expression of GADD proteins decreased with the Clark level of melanoma thickness. The survival time of patients correlated negatively with p53 positivity and positively with GADD 45 and 153 expression. Prognostic significance for GADD 34 was not found. Our conclusion is that GADD proteins play an important role in the malignant transformation of naevus to melanoma and GADD 45 and GADD 153 proteins can influence patient survival.
