一株新的急性髓系白血病细胞系SH-2的建立及其鉴定 |
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引用本文: | 仇惠英,孙爱宁,吴德沛,薛永权,张俊,戴海萍,潘金兰,吴亚芳,陈苏宁,王勇,沈娟. 一株新的急性髓系白血病细胞系SH-2的建立及其鉴定[J]. 中华血液学杂志, 2007, 30(1): 458-463. DOI: 10.3760/cma.j.issn.0253-2727.2009.07.009 |
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作者姓名: | 仇惠英 孙爱宁 吴德沛 薛永权 张俊 戴海萍 潘金兰 吴亚芳 陈苏宁 王勇 沈娟 |
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作者单位: | 苏州大学附属第一医院血液科、江苏省血液研究所,215006; |
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基金项目: | 江苏省自然科学基金卫生部基金 |
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摘 要: | Objective To establish and characterize a novel human myeloid leukemia cell line SH-2. Methods Bone marrow mononuclear cells(BMMNC) isolated from a AML-M2 patient, who failed to ob-tain complete remission after chemotherapy and allogenic bone marrow transplantation were passed in a long term IMDM culture medium supplemented with 20% fetal calf serum. Stromal cells were retained and rh-IL-3was added in the culture system. A new human myeloid leukemia cell line SH-2 was successfully established with a cytogeuetic characteristics of a loss of Y chromosome(- Y), a derivative chromosome 16 resulting from unbalanced translocation between chromosome 16 and 17, monosome 17, trisomy 19 and p53 alteration. Vari-ous methods were employed to characterize SH-2 cell line. Results SH-2 cells has been maintained without cytokine and stromal cells for more than 3 years without EB virus and mycoplasma contamination. SH-2 cells had the basically same morphological, immunophenotypic and cytogenetic features as the patient' s leukemia cells did, such as myeloid morphology, an immunophenotype of CD13+ , CD33+ , CD56+ , CD16/56+ and a hypodiploid karyotype of 45, X, - Y, der(16) t(16;17) (q24;q12) , - 17, + 19, which were gradually de-creased and replaced by the near-tetraploid cells with a karyotype of 73 - 102 (80), XX, - Y, - Y, del (lq31) ×2, der(16)t(16;17) (q24;q12) ×2, - 17, - 17, + 19, + 19. FISH and multiple FISH delineated all the abnormalities and revealed a loss of one p53 allele due to monosomy 17. DNA direct sequencing detec-ted a point mutation of CAG to CAT at codon 576 of exon 5 in another p53 allele. RT-PCR showed that SH-2 cells expressed apoptosis-related genes (bcl-2, Fas, GST- π and p21) rather than MDR-related genes. Short tandem repeat PCR provided powerful evidence for the derivation of SH-2 cell line from the patient' s leukemia cells. SH-2 cells had certain colony formation and tumorigenic capacities in nude and SCID mice. Conclu-sion SH-2 is a new myeloid leukemia cell line with a unique biology background, and will provide a useful tool for leukemia research.
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关 键 词: | 间质细胞 白细胞介素3 白血病,非淋巴细胞,急性 细胞系 |
Establishment and characterization of a new human myeloid leukemia cell line SH-2 |
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Abstract: | |
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Keywords: | Stromal cellslnterleukin-3Leukemia noalymphooytic acuteCell line |
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