Alteration of Serum Malondialdehyde Level As Biomarker of Oxidative Stress during Acute Myeloid Leukemia Treatment
Authors: Hlavackova Alzbeta , Vydra Jan , Chrastinova Leona , Kotlin Roman , Stikarova Jana , Suttnar Jiri , Dyr Jan E. 
Published: November 2019

Introduction
Oxidative stress as a disruption of redox homeostasis is a state where reactive oxygen species (ROS) concentration is transiently/chronically increased or ROS-scavening capacity is declined. ROS have an important role in intracellular signaling process. On the other side increased ROS production could lead to oxidative damage of biomolecules as lipids, DNA and proteins. Many types of cancer cells have elevated levels of ROS for promoting cell proliferation and differentiation [1,2]. In the context of hematopoietic malignancy, in acute myeloid leukemia (AML) there is evidence of elevated ROS and/or accompanying oxidative stress [3], however little is known about the changes of oxidative status during AML treatment.
The aim of this study was to estimate oxidative stress using the malondialdehyde (MDA) concentration as a marker of lipid peroxidation. The MDA concentrations were measured during treatment of AML patients in the initial stage (AML patients in diagnosis), after chemotherapy treatment (AML patients in remission), and after pretransplantation conditioning (AML patients before transplantation) in comparison with healthy controls.
Methods
Serum MDA concentration was analyzed in samples obtained at diagnosis, in remission, and before transplantation from patients (n=20) treated in the Institute of Hematology and Blood Transfusion, Prague, Czech Republic. As a control we used serum of healthy donors (n=13). Briefly, serum samples were prepared by alkaline hydrolysis, deproteinized and total MDA was derivatized by 2,4-dinitrophenylhydrazine (DNPH). MDA-DNPH derivates were separated by HPLC and determined using positive ion multiple reaction monitoring (MRM) mode by MS/MS.
Results
Using analysis of variance (ANOVA) we found significant differences in MDA concentrations in serum of AML patients and healthy controls (P < 0.0001). Moreover, Tukey post-hoc test applied to comparison of all treatment periods of AML patients with healthy controls revealed significantly increased MDA levels in the period before transplantation (P < 0.001) with respect to healthy controls. The MDA levels in the initial stage was also elevated as compared with healthy controls, but non significantly (P = 0.062). Comparing only AML patients in the individual treatment periods, we found significantly increased concentrations of MDA in samples of AML patients before transplantation with respect to AML patients in diagnosis (P = 0.023) and also to the patients in remission (P = 0.012).
Conclusion
Overall, throughout treatment periods for AML patients compared to healthy controls significant changes in MDA concentrations were identified. It is in agreement with previous findings stated that cancer cells have elevated levels of ROS possibly playing an important role in the initiation and progression of cancer. The highest MDA concentration was detected in samples of AML patients before transplantation which could be related to conditioning regimens [4]. The monitoring of AML patients at different times of disease treatment revealed that oxidative stress is changing not only as a result of disease progression but also as a result of treatment.
Acknowledgement
This work was supported by the European Regional Development Fund and the state budget of the Czech Republic (project AIIHHP: CZ.02.1.01/0.0/0.0/16_025/0007428, OP RDE, Ministry of Education, Youth and Sports), by the project of the Ministry of Health, Czech Republic, 00023736, by Grant from the Czech Science Foundation, Czech Republic, 19-02739S, and by ERDF OPPK CZ.2.16/3.1.00/24001.
[1] Boonstra J, Post JA. Gene 2004;337, 1-13.
[2] Schafer FQ, Buettner GR. Free Radic Biol. Med. 2001;30, 1191-1212.
[3] Hole PS, Zabkiewicz J, Munje C, Newton Z, Pearn L, White P, Marquez N, Hills RK, Burnett AK, Tonks A, Darley RL. Blood. 2013;122(19):3322-3330.
[4] Trachootham D, Alexandre J, Huang P. Nat Rev Drug Discov. 2009;8(7):579-591.

Disclosures
No relevant conflicts of interest to declare.