Peroxiredoxin-2 / PRDX2 is involved in redox regulation of the cell. It reduces peroxides with reducing equivalents provided through the thioredoxin system. Peroxiredoxin-2 / PRDX2 is not able to receive electrons from glutaredoxin. It may play an important role in eliminating peroxides generated during metabolism. Peroxiredoxin-2 / PRDX2 might participate in the signaling cascades of growth factors and tumor necrosis factor-alpha by regulating the intracellular concentrations of H2O2.
The pro-tumorigenic role of PRDX2 may be attributed to, at least partially, protecting cells from oxidative stress (3–6). Under oxidative stress, reactive oxygen species genways PRDX2 antibody also contribute to tumor necrosis factor-α (TNF-α)-induced cell death . However, the role of PRDX2 in TNF-α-induced cell death has not yet been established.

To determine whether other PRDX family members also bind to HIF-1α, we performed immunoprecipitation assays using whole cell lysates prepared from transfected HeLa cells that were exposed to hypoxia. As shown in Figure 1C, PRDX1-V5 strongly interacted with endogenous HIF-1α, similar to PRDX2-V5, whereas PRDX4-V5 and PRDX6-V5 weakly interacted with endogenous HIF-1α. These data indicate that PRDX1, PRDX2, PRDX4, and PRDX6, but not PRDX3 or PRDX5, interact with HIF-1α when overexpressed in HeLa cells.
Cells were then exposed to Celastrol for 24 h and stained with FITC-conjugated Annexin V and PI. Data were collected and analyzed using FACSCalibur flow cytometer. Phase contrast images of cells treated as indicated above were also captured using Nikon epifluorescence microscope . In addition, fluorescence images were captured following staining of cells with PI.

Three α subunits (HIF-1α, HIF-2α, and HIF-3α) and one β subunit (HIF-1β) have been identified [2-7]. Understanding the regulatory mechanisms underlying HIF transcriptional activity may yield novel therapeutic targets. Taken together, our results demonstrate that inhibiting PRDX2 expression promotes 5-FU-induced apoptosis in colon cancer cells via the PI3K/AKT signal pathway. PRDX2 may play a different role in the cell cycle of CRC tissues versus normal tissues. In bone marrow cells, a small-interfering RNA targeting PRDX2 caused G1-phase arrest in cells isolated from BALB/c mice . Additionally, depletion of PRDX2 has been shown to impair G1-phase progression and to inhibit cell proliferation in human keratinocytes cells .
As of January 1, 2022, Oncotarget has shifted to a continuous publishing model. Papers will now be published continuously within yearly volumes in their final and complete form and then quickly released to Pubmed. Human NPC cell line CNE2, obtained from Sun Yat-sen University Cancer Center, was cultured in RPMI-1640 medium plus 10% fetal bovine serum. We classified tumors with AJCC stage I + II as early-stage NPC as reported previously . Details for blood sample collection, processing and storage of serum sample of all participants were described in our previous publication . 16]; hence, the glutathione system may not be involved in regenerating native PRDX2.

The entire reaction mixture of each assay point was dispensed into 10 wells of 96-well plates (200 μl/well), and absorbance was read at 340 nm with a Biotek EL 808 Ultramicroplate Reader (). Control samples comprised a mixture of PRM-A and PRM-B prepared as already described but without H2O2 in PRM-A. The difference in NADPH content observed between the control and the reaction samples after 5-min incubation was considered the amount of NADPH lost due to peroxidase activity. The effects of carmustine [1,3-Bis(2-chloroethyl)-1-nitrosourea ] and diethyl maleate were investigated by mixing these drugs with the extracts, vortexing briefly, and incubating for 10 min before adding H2O2 and PRM-B.
Thus, the mammalian expression vector encoding GFP-PRDX2 and PRDX2 siRNA are useful tools to explore the role of PRDX2 in oxidative stress-mediated cell death in HCC SMMC-7721 cells. TNF-α usually does not induce cell death unless de novo protein synthesis is blocked . In this regard, protein synthesis inhibitor cloheximide is widely used to facilitate TNF-α-induced cell death, which includes both apoptosis and necrosis .
Anti-PRDX2 mouse monoclonal antibody was purchased from Abcam (). It was produced against recombinant human protein purified from Escherichia coli. A full-length N-terminal His-tagged recombinant human PRDX2 protein was procured from US Biological (). Anti-β-tubulin antibody E7 was purchased from Developmental Studies Hybridoma Bank (dshb.biology.uiowa.edu). Secondary antibodies conjugated to fluorescein isothiocyanate , tetramethyl rhodamine isothiocyanate , and horseradish peroxidase were procured from commercial sources (Zymed Lab Inc. []). Other reagents were purchased from Sigma () unless otherwise specified.

We added bio-Celastrol to HuProt protein array (Figure 3A & B) and identified the top-ranking proteins as antioxidant peroxiredoxins . All Prdx 1-4 interacted with Celastrol, with Prdx2 generating the greatest signal-to-noise ratio . Furthermore, assessment of other redox proteins showed low binding and underscored Prdx2 as the potential target of Celastrol . Interestingly, stable isotope labeling with amino acids in cell culture coupled to nanoLC-MS/MS analysis has recently shown Celastrol binding to Prdx1 in lymphoblastoid cell line and human colon cancer line HCT116 .
Thirty microliters of protein G agarose beads was added to the mixture and was rocked for an additional hour. Antibody control was prepared by mixing 3 μl of antibody with 400 μl of PBS extraction buffer and incubating with 30 μl of protein G agarose beads. After incubation, the beads were washed four times with PBS extraction buffer and finally were boiled with 30 μl of 2× concentrate loading buffer. In the present study, we investigated the presence of PRDX2 in spermatozoa and spermatids of mice and boar, its subcellular localization, and the peroxidase enzymatic activity of the sperm extracts. Primary antibodies like Anti-PRX II Antibody (A-2) for mammalian target proteins are recommended for the detection of a range of mammalian species, primarily of mouse, rat and human species.