Figure 1.
Increased oxidative stress in BCP‐ALL patients is accompanied by upregulation of TXN system antioxidant enzymes in BCP‐ALL primary cells and BCP‐ALL cell lines. (A) Redox biomarkers: ROM, BAP, and total thiol levels, as determined by free SH groups in proteins (SHP), were evaluated in the sera collected at diagnosis of BCP‐ALL pediatric (n = 35) and adult (n = 17) patients and compared with age‐related healthy subjects (HS, n = 21 and n = 14 for pediatric and adult HS, respectively). P values are presented for groups with significant differences (P < 0.05 in Mann–Whitney's U‐test). (B) mRNA levels of PRDX1, TXN1, and TXNRD1 in primary BCP‐ALL blasts (in total n = 129 pediatric ALL for PRDX1, n = 115 pediatric ALL for TXN1, TXNRD1, and n = 11 adult ALL for all genes) and in normal CD34+ (total RNA from CD34+ cells isolated from peripheral blood of multiple donors, two independent samples) and normal CD19+ (isolated from peripheral blood of three donors) cells. mRNA expression of the TXN system antioxidant enzymes was assessed by qPCR normalized to a reference gene, 60S ribosomal protein L29 (RPL29), as . Data are presented as individual values with mean ± SD. P values for comparisons between BCP‐ALL patients’ groups and controls are presented (Mann–Whitney's U‐test). (C) mRNA levels of PRDX1, TXN1, and TXNRD1 in BCP‐ALL cell lines were measured with qPCR and normalized to the housekeeping genes, RPL29 and/or β2‐microglobulin (B2M). Data show mean values from at least two independent experiments + SEM and are compared with control peripheral blood (PB) B cells and tonsillar germinal center (GC) B cells isolated from two healthy donors. *P < 0.05, ***P < 0.0001 by ANOVA with Dunnett's post hoc test (compared with GC B cells). (D) Representative western blot for the protein levels of TXN antioxidant enzymes in malignant B‐cell lines and normal GC B cells. Relative intensity to α‐tubulin calculated by densitometry (image lab Software; Bio‐Rad Laboratories) is presented under each band.