GRECAN, Centre François Baclesse, Université de Caen Basse-Normandie, Caen Cedex, France.
Leuk Res. 2010 Jun;34(6):800-8. doi: 10.1016/j.leukres.2009.10.018. Epub 2009 Nov 26.
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.
ZAP-70 表达是慢性淋巴细胞白血病的一个强有力的预后指标。然而，流式细胞术检测 ZAP-70 的定量分析缺乏足够的标准化。基于 ZAP-70 表达与其基因甲基化状态的相关性，我们开发了一种用于常规检测 ZAP-70 甲基化的定量焦磷酸测序法。ZAP-70 第一内含子中的四个 CpG 对（C-223、C-243、C-254 和 C-267）的甲基化与 ZAP-70 的抑制有关。此外，它与 CD38 表达（n=111，p<.0001）、IgHv 突变状态（n=106，p<.0001）、治疗时间（p<.0001）和总生存时间（p=.0014）相关。ZAP-70 的焦磷酸测序为流式细胞术提供了一个很好的替代方法。
