Cytological and spectroscopic characteristics of c-KIT N822K mutation in core binding factor acute myeloid leukemia cells.,Journal of Biophotonics

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Cytological and spectroscopic characteristics of c-KIT N822K mutation in core binding factor acute myeloid leukemia cells.
Journal of Biophotonics ( IF 2.0 ) Pub Date : 2020-06-05 , DOI: 10.1002/jbio.202000103
Yang Chen ₁ , Lingyan Wang ₂ , Xindi Lin ₁ , Qian Zhang ₁ , Yunchao Xu ₃ , Donghong Lin ₁ , Jianping Xu ₁ , Shangyuan Feng ₃ , Jianda Hu _{1,

2}

Affiliation

The frequency of N822K mutation is high in the A‐loop region of c‐KIT which is highly associated with poor prognosis of core binding factor acute myeloid leukemia. The current work used common assays including cell cycle, apoptosis, clone formation and western blot to perform cytological detection for HL60 (wild type), NB4 (carrying t[15;17] chromosome translocation) and Kasumi‐1 (with c‐KIT N822K mutation); and meanwhile, the laser tweezers Raman spectroscopy (LTRS) was also used to perform label‐free detection of single living cells. The results demonstrated that Kasumi‐1 cell line bearing c‐KIT N822K mutation has a stable cell cycle, while there was a significant difference between early and late apoptosis within 48 hours. The LTRS detection initially reflected the spectral differences induced by genetic abnormalities and highlighted progressive patterns of DNA and amino acids band contents which were appropriately consistent with that of cell clone ratio and the c‐KIT phosphorylation level. It is concluded that methodology of LTRS‐based single living cell characterization could be potential and effective to reveal gene mutation‐induced cell differentiation.

中文翻译：

核心结合因子急性髓系白血病细胞中c-KIT N822K突变的细胞学和光谱特征。

C-KIT A 环区 N822K 突变频率较高，与核心结合因子急性髓系白血病预后不良密切相关。目前的工作使用包括细胞周期、细胞凋亡、克隆形成和蛋白质印迹在内的常见检测方法对 HL60（野生型）、NB4（携带 t[15;17] 染色体易位）和 Kasumi-1（使用 c-KIT N822K）进行细胞学检测突变）; 同时，激光镊子拉曼光谱（LTRS）也被用于对单个活细胞进行无标记检测。结果表明携带c-KIT N822K突变的Kasumi-1细胞系具有稳定的细胞周期，而48小时内的早期和晚期细胞凋亡存在显着差异。LTRS 检测最初反映了由遗传异常引起的光谱差异，并突出了 DNA 和氨基酸条带含量的渐进模式，这些模式与细胞克隆率和 c-KIT 磷酸化水平适当一致。得出的结论是，基于 LTRS 的单个活细胞表征方法可能是潜在且有效地揭示基因突变诱导的细胞分化。

更新日期：2020-06-05

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