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Among the regions identified by the international research team are two, near the genes ARL15 and RREB1, that also show strong links to elevated levels of insulin and glucose in the body - two key characteristics of type 2 diabetes.
Per NeoGenomics Laboratories recommendations, (16) FISH was considered abnormal if at least one of the following criteria was met: (1) more than 16% of the nuclei showed more than 2 signals for RREB1 (red probe), (2) the ratio of red signal (RREB1) to aqua signal (centromere 6) greater than 1 was observed in more than 53% of the nuclei, (3) the ratio of yellow signal (MYB) to aqua signal (centromere 6) less than 1 was observed in more than 42% of the nuclei, and (4) more than 19% of the nuclei showed more than 2 signals for CCND1 (green probe).
MYB1 loss appears particularly common in spitzoid neoplasms and is rare in dysplastic nevi, whereas the opposite seems to be true for RREB1.
We suspect that this may be due in part to lower signal cutoffs for RREB1 (19%, criterion 1) and CCND1 (16%, criterion 2) used for MelanoSITE compared with those used by Gerami et al (6,17) (29% and 38%, respectively).
A commercially available FISH panel for the diagnosis of melanoma contains 4 probes directed against 2 chromosomes (6p25 RREB1, 6q23 MYB, 6centromere, 11q13 CCND1).
Sensitivity of fluorescence in situ hybridization for melanoma diagnosis using RREB1, MYB, Cep6, and 11q13 probes in melanoma subtypes.
From this analysis, the following 4 FISH criteria were identified: (1) if more than 38% of enumerated cells contained more than 2 signals for CCND1 (11q13) or (2) if more than 55% of nuclei contained more signals for 6p25 than for centromere 6 or (3) if more than 40% of nuclei contained fewer signals for MYB (6q23) than for centromere 6 or (4) if more than 29% of cells had more than 2 RREB1 (6p25) signals, then a case would be considered as positive for melanoma (18) (Table).
Abbott Molecular's 4-Probe Melanoma Fluorescence In Situ Hybridization Test Criterion Signal UCSF/Northwestern NeoGenomics University Cutoff, % Cutoff, % 6p25 gain > 2 RREB1 (red) > 29 > 16 6p25 gain RREB1 (red) > > 55 > 53 CEN6 (aqua) 6p23 loss MYB (yellow) < > 40 > 42 CEN6 (aqua) 11q13 gain > 2 CCND1 (green) > 38 > 19 Criteria and signal cutoffs were established by University of California in San Francisco UCSF/Northwestern University (Chicago, Illinois) studies and NeoGenomics Laboratories (Irvine, California).
In the FISH assay targeting loci RREB1 (6p25), MYB (6q23), CCND1 (11q13), and CEP (centromere of chromosome 6), 15 of 16 positive cases demonstrated aberrations in chromosome 6 and one case showed gains in 11q13 (CCND1) in their control group of 17 lentiginous nevi, FISH evaluation was negative.
The assay Vysis LSI RREB1/LSI MYB/LSI CCND1/CEP 6 probes, is designed to detect copy number of RREB1 (6p25; red spectrum), MYB (6q23; gold spectrum) and CCND1 (11q13; green spectrum) genes and of centromere 6 (cep6; aqua spectrum) by using FISH in formalin-fixed, paraffin-embedded skin tissue sections (Figure 1).
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- RREP Without RREQ