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首页> 外文期刊>Genes, Chromosomes and Cancer >Molecular cytogenetics of the acute promyelocytic leukemia-derived cell line NB4 and of four all-trans retinoic acid-resistant subclones.
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Molecular cytogenetics of the acute promyelocytic leukemia-derived cell line NB4 and of four all-trans retinoic acid-resistant subclones.

机译：急性早幼粒细胞白血病衍生细胞系NB4和四个全反式维甲酸抗性亚克隆的分子细胞遗传学。

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The retinoic acid (RA)-sensitive NB4 cell line was the first established acute promyelocytic leukemia (APL) permanent cell line. It harbors the (15;17) translocation, which fuses the PML and RARA genes. Given the low frequency of APLs, their generally low white blood cell count, and the difficulty to work on APL patient cells, this cell line represents a remarkable tool for biomolecular studies. To investigate possible mechanisms of retinoid resistance, subclones of NB4 resistant to all-trans retinoic acid (ATRA) were established. To characterize better the parental NB4 cell line and four ATRA-resistant subclones (NB4-R4, NB4-A1, NB4-B1, and NB4-007/6), we have performed both conventional and 24-color FISH karyotyping. Thus, we could identify all chromosomal abnormalities including marker chromosomes that were unclassified with R banding. Moreover, we have performed dual-color FISH by use of specific PML and RARA probes, to evaluate the number of copies for each gene and fusion gene. Interestingly, the number of copies of PML, RARA, and fusion genes was different for each cell line. Finally, we assessed the presence of the PML, RARA, PML/RARA, and RARA/PML transcripts by RT-PCR and of the PML/RARA and RARA proteins by Western blotting in all the cell lines. These data could focus further research for a better understanding of the molecular mechanisms underlying response or resistance to differentiating and/or apoptotic reagents.
机译：视黄酸（RA）敏感的NB4细胞系是最早建立的急性早幼粒细胞白血病（APL）永久细胞系。它具有（15; 17）易位，融合了PML和RARA基因。鉴于APL的频率低，白细胞数量普遍较低以及难以处理APL患者细胞，该细胞系代表了生物分子研究的重要工具。为了研究类维生素A耐药的可能机制，建立了对全反式维甲酸（ATRA）耐药的NB4亚克隆。为了更好地表征亲本NB4细胞系和四个ATRA抗性亚克隆（NB4-R4，NB4-A1，NB4-B1和NB4-007 / 6），我们已经进行了常规和24色FISH核型分析。因此，我们可以识别所有染色体异常，包括未用R条带分类的标记染色体。此外，我们已经通过使用特定的PML和RARA探针进行了双色FISH，以评估每个基因和融合基因的拷贝数。有趣的是，每种细胞系的PML，RARA和融合基因的拷贝数都不同。最后，我们通过RT-PCR评估了PML，RARA，PML / RARA和RARA / PML转录本的存在，并通过Western印迹评估了所有细胞系中PML / RARA和RARA蛋白的存在。这些数据可能会集中于进一步的研究，以更好地理解对分化和/或凋亡试剂的反应或抗性的分子机制。

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