Jetly Sunita, Verma Neha, Naidu Kumar, Faiq Muneeb Ahmad, Seth Tulika, Saluja Daman
Associate Professor, Department of Biotechnology, Dr. B. R. Ambedkar Center for Biomedical Research, University of Delhi, New Delhi, India.
Project Fellow, Department of Biotechnology, Dr. B. R. Ambedkar Center for Biomedical Research, University of Delhi, New Delhi, India.
J Clin Diagn Res. 2017 Aug;11(8):XC01-XC05. doi: 10.7860/JCDR/2017/28565.10425. Epub 2017 Aug 1.
There is a significant difference in the Reactive Oxygen Species (ROS) levels of Chronic Myeloid Leukaemia (CML) patients before and during treatment with Tyrosine Kinase Inhibitors (TKIs). This is because high ROS levels support oncogenic phenotype of CML by inducing proliferation pathway and accumulation of further genetic mutations. Often the measurement is done on WBC or serum for ascertaining one type of ROS species, but measurement of global ROS in fresh whole blood will give more accurate estimation of ROS.
To measure global ROS in peripheral blood of CML patients.
A case control study was undertaken to measure ROS in peripheral blood of CML patients from Northern India. CML patients on TKIs (n=40 on imatinib herein called treated) and untreated (n=17, who were not on any TKIs or alternative medicine, called as treatment naive) and 52 healthy controls were also enrolled. Chemiluminescent assay was carried out using luminol as signal enhancer in 400 µl of blood to measure ROS. The chemiluminescence was measured as Relative Light Units (RLU)/sec/104 WBC. Data was presented in terms of mean±SE or geometric mean (95% Confidence Interval) for continuous variables and percentage for categorical variables. Groups were compared using two sample t-test for continuous variables and chi-square test for categorical variables.
The WBC profile and ROS levels of patients taking TKIs were quite similar and showed no significant difference (p<0.999) compared to healthy controls. In contrast, significant increase was observed in the ROS levels of CML patients not on TKIs (untreated) compared to patients under treatment (p<0.029) and healthy controls (p<0.007). We also observed that the absolute ROS values and WBC counts were higher in untreated patients compared to patients on TKIs and healthy controls, even though mean ROS value was less.
To ascertain the alterations in ROS levels of CML patients before and during treatment with TKIs, it is better to measure global ROS in fresh whole blood by chemiluminescent method using luminol. Luminol assay is a quick, easy and inexpensive method to measure global ROS. Patient under treatment with TKIs show significant decrease in ROS levels almost similar to the levels measured in healthy controls yet the mechanisms by which this decrease occurs needs to be elucidated.
慢性髓性白血病（CML）患者在接受酪氨酸激酶抑制剂（TKIs）治疗之前和治疗期间的活性氧（ROS）水平存在显著差异。这是因为高ROS水平通过诱导增殖途径和进一步的基因突变积累来支持CML的致癌表型。通常是在白细胞或血清上进行测量以确定一种类型的ROS，但在新鲜全血中测量整体ROS将能更准确地估计ROS。
测量CML患者外周血中的整体ROS。
进行了一项病例对照研究，以测量来自印度北部的CML患者外周血中的ROS。还纳入了接受TKIs治疗的CML患者（此处伊马替尼治疗组n = 40，称为治疗组）和未治疗患者（n = 17，未接受任何TKIs或替代药物治疗，称为初治组）以及52名健康对照者。使用鲁米诺作为信号增强剂，在400微升血液中进行化学发光测定以测量ROS。化学发光以相对光单位（RLU）/秒/10⁴白细胞来衡量。连续变量的数据以均值±标准误或几何均值（95%置信区间）表示，分类变量的数据以百分比表示。连续变量使用两样本t检验进行组间比较，分类变量使用卡方检验进行组间比较。
接受TKIs治疗的患者的白细胞谱和ROS水平非常相似，与健康对照相比无显著差异（p < 0.999）。相比之下，未接受TKIs治疗（未治疗）的CML患者的ROS水平与接受治疗的患者相比显著升高（p < 0.029），与健康对照相比也显著升高（p < 0.007）。我们还观察到，未治疗患者的绝对ROS值和白细胞计数高于接受TKIs治疗的患者和健康对照，尽管平均ROS值较低。
为了确定CML患者在接受TKIs治疗之前和治疗期间ROS水平的变化，最好通过使用鲁米诺的化学发光法在新鲜全血中测量整体ROS。鲁米诺测定法是一种快速、简便且廉价的测量整体ROS的方法。接受TKIs治疗的患者的ROS水平显著降低，几乎与健康对照者测量的水平相似，但这种降低发生的机制仍需阐明。
