Sotlar K, Marafioti T, Griesser H, Theil J, Aepinus C, Jaussi R, Stein H, Valent P, Horny H P
Institute of Pathology, University Hospital Tübingen, Germany.
Mol Pathol. 2000 Aug;53(4):188-93. doi: 10.1136/mp.53.4.188.
BACKGROUND/AIMS: The occurrence of myeloid leukaemia in patients with systemic mastocytosis is a well recognised phenomenon. However, the pathophysiological basis of such a coevolution has not been clarified. Recent data have shown that the c-kit mutation Asp 816 to Val is detectable in neoplastic mast cells in most patients with systemic mastocytosis, including those who have associated haematological disorders. The aim of this study was to study clonal disease evolution by analysing bone marrow cells from a patient with systemic mastocytosis and associated chronic myelomonocytic leukaemia (CMML) for the presence of this mutation.
The DNA of microdissected bone marrow cells from a patient with systemic mastocytosis and associated CMML was analysed for the presence of the c-kit mutation Asp 816 to Val by means of HinfI digestion and direct sequencing of semi-nested polymerase chain reaction (PCR) products.
The two neoplasms could easily be identified and discriminated in paraffin wax embedded bone marrow sections by tryptase and chloroacetate esterase staining. A total number of 10 tryptase positive systemic mastocytosis infiltrates and 10 tryptase negative CMML infiltrates were removed by microdissection. As assessed by HinfI digestion and direct sequencing of semi-nested PCR products, the c-kit mutation Asp 816 to Val was detected in five of seven systemic mastocytosis infiltrates and four of six CMML infiltrates. By contrast, no c-kit mutation Asp 816 to Val was found in bone marrow infiltrates in patients with CMML without associated systemic mastocytosis (n = 20).
These data support a monoclonal evolution of systemic mastocytosis and concurrent CMML in the patient studied.
背景/目的：系统性肥大细胞增多症患者发生髓系白血病是一种已被充分认识的现象。然而，这种共同演变的病理生理基础尚未阐明。最近的数据表明，在大多数系统性肥大细胞增多症患者的肿瘤性肥大细胞中可检测到c-kit基因第816位密码子由天冬氨酸突变为缬氨酸，包括那些伴有血液系统疾病的患者。本研究的目的是通过分析一名系统性肥大细胞增多症合并慢性粒-单核细胞白血病（CMML）患者的骨髓细胞中该突变的存在情况，来研究克隆性疾病的演变。
通过HinfI酶切和半巢式聚合酶链反应（PCR）产物的直接测序，分析一名系统性肥大细胞增多症合并CMML患者经显微切割的骨髓细胞DNA中c-kit基因第816位密码子由天冬氨酸突变为缬氨酸的情况。
通过胰蛋白酶和氯乙酸酯酶染色，在石蜡包埋的骨髓切片中可以很容易地识别和区分这两种肿瘤。通过显微切割去除了总共10个胰蛋白酶阳性的系统性肥大细胞增多症浸润灶和10个胰蛋白酶阴性的CMML浸润灶。通过HinfI酶切和半巢式PCR产物的直接测序评估，在7个系统性肥大细胞增多症浸润灶中的5个和6个CMML浸润灶中的4个检测到了c-kit基因第816位密码子由天冬氨酸突变为缬氨酸。相比之下，在不伴有系统性肥大细胞增多症的CMML患者的骨髓浸润灶中未发现c-kit基因第816位密码子由天冬氨酸突变为缬氨酸（n = 20）。
这些数据支持了所研究患者中系统性肥大细胞增多症和并发CMML的单克隆演变。
