Document Detail


Quantitative relationships between pigment-related mRNA and biochemical melanoma markers in melanoma cell lines.
MedLine Citation:
PMID:  12140375     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The use of reverse transcription polymerase chain reaction (RT-PCR) analysis of melanoma-specific transcripts for the identification of circulating melanoma cells has shown very variable results in different studies on melanoma patients. We have therefore developed quantitative methods to study both analytical and biological variations as possible causes of this phenomenon. Pigment-related and S-100 beta transcripts were quantified in 12 different melanoma cell lines and related to the amounts of 5-S-cysteinyldopa, pigment and S-100B protein. A real-time PCR method was used and the results were expressed as absolute number of transcripts per cell. Tyrosinase, tyrosinase-related protein (TRP)-1, TRP-2 and MART-1/Melan-A mRNA varied from undetectable (< 10(-4) transcripts/cell) to 10(3) transcripts/cell, i.e. by a factor > 10(7) in the different cell lines. S-100 beta mRNA varied from 2.8 to 165 transcripts/cell, i.e. by a factor of 60. Tyrosinase, TRP-1 and TRP-2 mRNA correlated significantly with the amount of 5-S-cysteinyldopa, an intermediate pigment metabolite (P < 0.001, P < 0.001 and P < 0.01, respectively). The amount of S-100 beta mRNA correlated significantly with the amount of S-100B protein (P < 0.001). No cross-correlations were seen between the pigment-related and S-100-related analytes. We conclude that one reason behind the negative results of RT-PCR measurement of pigment-related mRNA may be that these transcripts are not always expressed in the particular cells present in the patient's blood. Furthermore, variation in the expression of the order of 10(7) must have great impact on the diagnostic sensitivity. Measurement of S-100 beta mRNA would be more sensitive, but the use of this transcript is hampered by its presence in the blood cells.
Authors:
M Johansson; A Takasaki; L Lenner; K Arstrand; B K?gedal
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Melanoma research     Volume:  12     ISSN:  0960-8931     ISO Abbreviation:  Melanoma Res.     Publication Date:  2002 Jun 
Date Detail:
Created Date:  2002-07-25     Completed Date:  2003-01-30     Revised Date:  2010-06-10    
Medline Journal Info:
Nlm Unique ID:  9109623     Medline TA:  Melanoma Res     Country:  England    
Other Details:
Languages:  eng     Pagination:  193-200     Citation Subset:  IM    
Affiliation:
Department of Biomedicine and Surgery, Division of Clinical Chemistry, Link?ping University, S-581 85 Link?ping, Sweden.
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MeSH Terms
Descriptor/Qualifier:
Antigens, Neoplasm
Gene Expression Profiling
Humans
Intramolecular Oxidoreductases / genetics
Melanins / analysis,  biosynthesis
Melanoma / blood,  chemistry,  genetics,  pathology*
Membrane Glycoproteins*
Monophenol Monooxygenase / genetics
Neoplasm Proteins / genetics
Neoplastic Cells, Circulating
Nerve Growth Factors
Oxidoreductases*
Pigmentation
Polymerase Chain Reaction
Proteins / genetics
Pyrroles / analysis
RNA, Messenger / analysis*
RNA, Neoplasm / analysis*
S100 Proteins / analysis,  genetics
Tumor Cells, Cultured / chemistry
Tumor Markers, Biological / analysis*
Tyrosine / analogs & derivatives*,  analysis
Chemical
Reg. No./Substance:
0/Antigens, Neoplasm; 0/MART1 antigen; 0/Melanins; 0/Membrane Glycoproteins; 0/Neoplasm Proteins; 0/Nerve Growth Factors; 0/Proteins; 0/Pyrroles; 0/RNA, Messenger; 0/RNA, Neoplasm; 0/S-100 calcium-binding protein beta subunit; 0/S100 Proteins; 0/Tumor Markers, Biological; 55520-40-6/Tyrosine; 74923-08-3/4-amino-3-hydroxyphenylalanine; 945-32-4/pyrrole-2,3,5-tricarboxylic acid; EC 1.-/Oxidoreductases; EC 1.14.18.-/TYRP1 protein, human; EC 1.14.18.-/tyrosinase-related protein-1; EC 1.14.18.1/Monophenol Monooxygenase; EC 5.3.-/Intramolecular Oxidoreductases; EC 5.3.3.12/dopachrome isomerase

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