ZAP-70 intron1 DNA methylation status: determination by pyrosequencing in B chronic lymphocytic leukemia.
ZAP-70 expression is a strong prognostic indicator in chronic lymphocytic leukemia. However, ZAP-70 quantification by flow cytometry lacks sufficient standardization. Based upon the correlation between ZAP-70 expression and its gene methylation status, we have developed a quantitative pyrosequencing assay for the determination of ZAP-70 methylation adapted for routine use. Methylation in four CpG pairs (C-223, C-243, C-254, and C-267) in the first intron of ZAP-70 is associated with repression of ZAP-70. Moreover, it correlates with CD38 expression (n=111, p<.0001), IgHv mutation status (n=106, p<.0001), time to treatment (p<.0001), and overall survival (p=.0014). Pyrosequencing of ZAP-70 provides a good alternative to flow cytometry.
Chantepie
Sylvain P
SP
GRECAN, Centre François Baclesse, Université de Caen Basse-Normandie, Caen Cedex, France. chantepie-s@chu-caen.fr
Vaur
Dominique
D
Grunau
Christoph
C
Salaün
Véronique
V
Briand
Mélanie
M
Parienti
Jean-Jacques
JJ
Heutte
Natacha
N
Cheze
Stéphane
S
Roussel
Mikel
M
Gauduchon
Pascal
P
Leporrier
Michel
M
Krieger
Sophie
S
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D019175:DNA Methylation; D018450:Disease Progression; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D015973:Gene Expression Regulation, Leukemic; D006801:Humans; D007438:Introns / Q000235:genetics; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D011237:Predictive Value of Tests; D011379:Prognosis; D017422:Sequence Analysis, DNA / Q000379:methods*; D051746:ZAP-70 Protein-Tyrosine Kinase / Q000235:genetics*
D051746:ZAP-70 Protein-Tyrosine Kinase; C497261:ZAP70 protein, human