M-FISH 用于骨髓增生异常综合征细胞株MUTZ-1复杂核型检测分析.docVIP

M-FISH 用于骨髓增生异常综合征细胞株MUTZ-1复杂核型检测分析 　 作者：陈宝安，夏国华，李建勇，肖冰，邵泽叶 陈宁娜，高冲，吴雨洁 【摘要】 本研究的目的是观察伴有5号染色体缺失的MDS 细胞株MUTZ-1细胞的遗传学变化。首先采用R显带技术对染色体标本进行核型分析，再以Vysis Spectra VysionTM M-FISH作为探针，检测并分析其复杂异常核型。结果表明： M-FISH分析显示有明显的高频率染色体的易位、插入、断裂与重接、缺失、数目增多；染色体分析揭示核型为50，xx，der(1) t(1；2)，ins(1；14)，+der(2)t(2；19)，der(3)t(3；5)，der(3)(3::5::22)，5q-，der(6)t(3；6)，der(7)(18::7::17)，+8，+der(9)t(1；9)，der(10)t(1；10)，+11，+12，der(?13)(10::13::5::8)，der(14)t(8；14)，der(14)t(14，15)，der(15)t(15；21)×2，+17，+18，-21，-22. 结论: MDS 细胞株MUTZ-1在M-FISH检测下有显著复杂的核型变化；M-FISH能增加高度复杂的异常染色体检测的准确性，有助于MDS的诊断和预后评估。 【关键词】 骨髓增生异常综合征 　　Cytogenetic studies play a major role in confirmation of diagnosis and prediction of clinical outcome in MDS，and have contributed to the understanding of its pathogenesis. No cytogenetic abnormality is specific for MDS or for a specific morphologic subgroup of MDS［1］. However，some unique cytogenetic/morphologic correlations exist，the most common of which is the “5q-syndrome”. In this report，we describe a myelodysplastic syndrome cell line (MUTZ-1) from a little girl diagnosed as having REAB with complex karyotype by metaphase fluorescence in situ hybridization (M-FISH). 　　Materials and Methods 　　Reagents and instruments 　　Vysis Spectra VysionTM M-FISH kit and DAPI II Counterstain (125 ng/ml in antifade mounting solution) were purchased from American Vysis Corporation. Rubber cement for sealing the coverslip came from American Starkey Chemical. RNase and pepsin were purchased from Sigama Corporation. In addition，The Leica DMRA2 fluorescence microscopy with a charge coupled device camera and Cytovision Specta Vysion software were respectively purchased from German Leica corporation，Applied Imaging corporation. 　　Case description 　　The patient was a 5-year-old girl who presented to hospital in 1997 with general fatigue. She was diagnosed as having REAB according to the FAB classification. Myelodysplastic syndrome cell line (MUTZ-1) was established and cultured at the time. The karyotype of the or