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首页> 外文期刊>Experimental Hematology: Official Publication of the International Society for Experimental Hematology >Establishment and characterization of a new human acute myelocytic leukemia cell line SH-2 with a loss of Y chromosome, a derivative chromosome 16 resulting from an unbalanced translocation between chromosomes 16 and 17, monosomy 17, trisomy 19, and p53 alteration.
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Establishment and characterization of a new human acute myelocytic leukemia cell line SH-2 with a loss of Y chromosome, a derivative chromosome 16 resulting from an unbalanced translocation between chromosomes 16 and 17, monosomy 17, trisomy 19, and p53 alteration.

机译：建立并鉴定了一种新的人类急性髓细胞白血病细胞系SH-2，该细胞系具有Y染色体缺失，16号染色体与17号染色​​体之间不平衡易位导致的衍生性16号染色体，17号染色​​体，19号染色体，以及p53突变。

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OBJECTIVE: To report here a new acute myelocytic leukemia (AML) cell line SH-2 and describe its biological characteristics. MATERIALS AND METHODS: Mononuclear cells isolated from a patient with AML-M2 subtype were passaged by liquid culture medium. Interleukin-3 and bone marrow stromal cells were used to support cell proliferation at the first 3 months. Various methods, including cytogenetic analysis, fluorescence in situ hybridization (FISH), multiplex FISH (M-FISH), reverse transcriptase polymerase chain reaction (RT-PCR), multiplex RT-PCR, short tandem repeat (STR)-PCR, direct sequencing of DNA, clonogenic assay, and tumorigenicity in nude and severe combined immunodeficient (SCID) mice were employed to identify and characterize SH-2 cell line. RESULTS: SH-2 cells were maintained without cytokine and stromal cells for 3 years. It had no Epstein-Barr virus or mycoplasma contamination. The SH-2 cell line showed typical myelocytic features in morphology and simultaneous strongly expressed myeloid antigens (CD13, 99.6% and CD33, 99.26%) and natural killer (NK)-related antigens (CD56, 99.5% and CD16/56, 99.62%) suggesting that SH-2 is an AML cell line with NK-antigen expression. SH-2 cell line initially showed a karyotype of 45, X, -Y, der(16)t(16;17)(q24;q12), -17, +19. During the passage period, the cells with a hypodiploid karyotype gradually decreased and were replaced by the near-tetraploid cells with a karyotype of 71-105(86), XX, -Y, -Y, der(16)t(16;17)x2, -17, -17, +19, +19. FISH and M-FISH delineated all abnormalities. SH-2 cells had the approximately same morphological, immunophenotypical, and cytogenetic features as the patient's leukemia cells had. STR-PCR provided powerful evidence for the derivation of SH-2 cell line from the patient's leukemia cells. SH-2 cells showed multiple drug resistance (MDR), which may be related to the p53 gene alteration, including the loss of one p53 allele due to the monosomy 17 and a point mutation of CAG to CAT at codon 576 of exon 5 in another p53 allele resulting in the loss of p53 gene function. In addition, SH-2 cell line did not express MDR-related genes, such as MDR1, multidrug resistance-related protein, and lung resistance protein, but expressed apoptosis-related genes, such as Bcl-2, Fas, glutathione S-transferase-pi, and p21, which were also related to the MDR. SH-2 cell line had tumorigenic capacities in nude and SCID mice. CONCLUSION: Because SH-2 cell line had a clear biology background, it will provide a useful tool for the study of the pathogenesis and treatment strategy of AML with MDR.
机译：目的：在此报告一种新的急性粒细胞白血病（AML）细胞系SH-2，并描述其生物学特性。材料与方法：从AML-M2亚型患者中分离出的单核细胞通过液体培养基传代。白细胞介素3和骨髓基质细胞在头3个月用于支持细胞增殖。多种方法，包括细胞遗传学分析，荧光原位杂交（FISH），多重FISH（M-FISH），逆转录酶聚合酶链反应（RT-PCR），多重RT-PCR，短串联重复（STR）-PCR，直接测序裸鼠和重度联合免疫缺陷（SCID）小鼠的DNA克隆测定，克隆形成测定和致瘤性被用于鉴定和表征SH-2细胞系。结果：SH-2细胞维持3年无细胞因子和基质细胞。它没有爱泼斯坦-巴尔病毒或支原体污染。 SH-2细胞系在形态上表现出典型的髓细胞特征，并同时强烈表达髓样抗原（CD13，99.6％和CD33，99.26％）和自然杀伤（NK）相关抗原（CD56，99.5％和CD16 / 56，99.62％）。 ）表明SH-2是具有NK抗原表达的AML细胞系。 SH-2细胞系最初显示出45，X，-Y，der（16）t（16; 17）（q24; q12），-17，+ 19的核型。在传代期间，具有二倍体核型的细胞逐渐减少，并被具有71-105（86），XX，-Y，-Y，der（16）t（16; 17）的核型的近四倍体细胞替代。 ）x2，-17，-17，+ 19，+ 19。 FISH和M-FISH描绘了所有异常情况。 SH-2细胞与患者的白血病细胞具有大致相同的形态，免疫表型和细胞遗传学特征。 STR-PCR为从患者的白血病细胞衍生SH-2细胞系提供了有力的证据。 SH-2细胞表现出多重耐药性（MDR），这可能与p53基因的改变有关，包括一个17号单体性导致一个p53等位基因的丢失，以及另一个外显子5密码子576处CAG突变为CAT。 p53等位基因导致p53基因功能丧失。此外，SH-2细胞系不表达MDR相关基因，例如MDR1，多药耐药相关蛋白和肺耐药蛋白，但是表达凋亡相关基因，例如Bcl-2，Fas，谷胱甘肽S-转移酶-pi和p21，它们也与MDR有关。 SH-2细胞系在裸鼠和SCID小鼠中具有致瘤能力。结论：SH-2细胞系具有明确的生物学背景，它将为研究MDR伴发AML的发病机制和治疗策略提供有用的工具。

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