Development of a Real-Time Reverse Transcription Polymerase Chain Reaction Assay for c-myc Expression That Allows the Identification of a Subset ... View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2003-02-01

AUTHORS

Ana-Isabel Sáez, María-Jesús Artiga, Cristina Romero, Sandra Rodríguez, Juan-Cruz Cigudosa, Alberto Pérez-Rosado, Isabel Fernández, Margarita Sánchez-Beato, Esther Sánchez, Manuela Mollejo, Miguel Á Piris

ABSTRACT

Absence of a reliable method for determining the level of c-myc expression has impeded the analysis of its biological and clinical relevance in tumors. We have standardized the conditions for a real-time reverse transcription polymerase chain reaction analysis for c-myc expression, including the selection of an endogenous reference (18S rRNA), the adequate number of measurements for each sample (2 cDNA in triplicate), and suitable controls for determining inter- and intrarun variability (standard curve and calibrator). Subsequently, in a series of 56 non-Hodgkin's lymphomas, we analyzed the expression of c-myc mRNA, using real-time reverse transcription polymerase chain reaction, and of other functionally related proteins (bcl-6, p27, cyclin D3, and p53). As expected, all eight Burkitt's lymphoma cases analyzed had high levels of c-myc mRNA expression compared with that observed in reactive lymphoid tissue. There was a wider range of expression in diffuse large B-cell lymphoma, with 30% (15 of 48) of cases overexpressing c-myc. This overexpression was largely independent of c-myc translocations (4 of 5), as demonstrated by fluorescence in situ hybridization. In this large B-cell lymphoma series, a high level of c-myc expression predicted lower survival probability, irrespectively of the International Prognostic Index risk group classification. A slightly increased frequency of p53 inactivation was observed in the cases with c-myc overexpression, which suggests a growth advantage in lymphomas with concurrent deregulation of c-myc and p53. In addition, a moderate increase in bcl-6 protein expression was observed in the c-myc–positive cases, suggesting the existence of a complex interrelationship between these two genes. These findings suggest that c-myc may play a relevant role in the pathogenesis of a subset of large B-cell lymphoma and suggest the existence of additional regulatory mechanisms of c-myc expression to c-myc rearrangements. More... »

PAGES

143-152

Identifiers

URI

http://scigraph.springernature.com/pub.10.1097/01.lab.0000057000.41585.fd

DOI

http://dx.doi.org/10.1097/01.lab.0000057000.41585.fd

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1013853338

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/12594230


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34 schema:description Absence of a reliable method for determining the level of c-myc expression has impeded the analysis of its biological and clinical relevance in tumors. We have standardized the conditions for a real-time reverse transcription polymerase chain reaction analysis for c-myc expression, including the selection of an endogenous reference (18S rRNA), the adequate number of measurements for each sample (2 cDNA in triplicate), and suitable controls for determining inter- and intrarun variability (standard curve and calibrator). Subsequently, in a series of 56 non-Hodgkin's lymphomas, we analyzed the expression of c-myc mRNA, using real-time reverse transcription polymerase chain reaction, and of other functionally related proteins (bcl-6, p27, cyclin D3, and p53). As expected, all eight Burkitt's lymphoma cases analyzed had high levels of c-myc mRNA expression compared with that observed in reactive lymphoid tissue. There was a wider range of expression in diffuse large B-cell lymphoma, with 30% (15 of 48) of cases overexpressing c-myc. This overexpression was largely independent of c-myc translocations (4 of 5), as demonstrated by fluorescence in situ hybridization. In this large B-cell lymphoma series, a high level of c-myc expression predicted lower survival probability, irrespectively of the International Prognostic Index risk group classification. A slightly increased frequency of p53 inactivation was observed in the cases with c-myc overexpression, which suggests a growth advantage in lymphomas with concurrent deregulation of c-myc and p53. In addition, a moderate increase in bcl-6 protein expression was observed in the c-myc–positive cases, suggesting the existence of a complex interrelationship between these two genes. These findings suggest that c-myc may play a relevant role in the pathogenesis of a subset of large B-cell lymphoma and suggest the existence of additional regulatory mechanisms of c-myc expression to c-myc rearrangements.
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42 BCL-6 protein expression
43 Burkitt lymphoma cases
44 Hodgkin's lymphoma
45 absence
46 addition
47 additional regulatory mechanisms
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49 advantages
50 analysis
51 assays
52 c-Myc
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57 c-myc rearrangement
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67 concurrent deregulation
68 conditions
69 control
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71 development
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73 endogenous reference
74 existence
75 expression
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77 fluorescence
78 frequency
79 genes
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83 hybridization
84 identification
85 inactivation
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87 interrelationships
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89 levels
90 lower survival probability
91 lymphoid tissue
92 lymphoma
93 lymphoma cases
94 mRNA
95 mRNA expression
96 measurements
97 mechanism
98 method
99 moderate increase
100 number
101 overexpression
102 p53
103 p53 inactivation
104 pathogenesis
105 polymerase chain reaction
106 polymerase chain reaction analysis
107 polymerase chain reaction assays
108 probability
109 protein
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111 range
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114 reaction assays
115 reactive lymphoid tissues
116 real-time reverse transcription polymerase chain reaction assays
117 real-time reverse transcription-polymerase chain reaction
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119 rearrangement
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128 risk group classification
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131 selection
132 series
133 situ hybridization
134 subset
135 suitable control
136 survival probability
137 tissue
138 transcription-polymerase chain reaction
139 transcription-polymerase chain reaction analysis
140 transcription-polymerase chain reaction assays
141 translocation
142 tumors
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