Targeting peroxiredoxin 1 impairs growth of breast cancer cells and potently sensitises these cells to prooxidant agents

Abstract

Background Our previous work has shown peroxiredoxin-1 (PRDX1), one of major antioxidant enzymes, to be a biomarker in human breast cancer. Hereby, we further investigate the role of PRDX1, compared to its close homolog PRDX2, in mammary malignant cells. Methods CRISPR/Cas9- or RNAi-based methods were used for genetic targeting PRDX1/2. Cell growth was assessed by crystal violet, EdU incorporation or colony formation assays. In vivo growth was assessed by a xenotransplantation model. Adenanthin was used to inhibit the thioredoxin-dependent antioxidant defense system. The prooxidant agents used were hydrogen peroxide, glucose oxidase and sodium L-ascorbate. A PY1 probe or HyPer-3 biosensor were used to detect hydrogen peroxide content in samples. Results PRDX1 downregulation significantly impaired the growth rate of MCF-7 and ZR-75-1 breast cancer cells. Likewise, xenotransplanted PRDX1 - deficient MCF-7 cells presented a retarded tumour growth. Furthermore, genetic targeting of PRDX1 or adenanthin, but not PRDX2, potently sensitised all six cancer cell lines studied, but not the non-cancerous cells, to glucose oxidase and ascorbate. Conclusions Our study pinpoints the dominant role for PRDX1 in management of exogeneous oxidative stress by breast cancer cells and substantiates further exploration of PRDX1 as a target in this disease, especially when combined with prooxidant agents.

Triple Combination of Ascorbate, Menadione and the Inhibition of Peroxiredoxin-1 Produces Synergistic Cytotoxic Effects in Triple-Negative Breast Cancer Cells

“…The cytotoxicity of the prooxidant compounds was assessed as described previously [29]. Briefly, cells were treated with increasing concentrations of Men, Asc, or Men/Asc combination, AUR alone, or combined with either Men or Asc for 24 h. Next, the crystal violet assay was applied to determine the viability of cultured cells [29].…”

“…The cytotoxicity of the prooxidant compounds was assessed as described previously [29]. Briefly, cells were treated with increasing concentrations of Men, Asc, or Men/Asc combination, AUR alone, or combined with either Men or Asc for 24 h. Next, the crystal violet assay was applied to determine the viability of cultured cells [29]. Drug combination studies and their synergy quantification were calculated using the Chou-Talalay method by CompuSyn software v1.0 (Combosyn, Inc., Paramus, NJ, USA) [30] as described previously [9].…”

“…Given the information that downregulation of PRDX1 potently sensitizes TNBC cells to either Asc [29] or Men (Figure 2), we evaluated the response of two TNBC cell lines, i.e., MDA-MB-231-…”

“…Given the information that downregulation of PRDX1 potently sensitizes TNBC cells to either Asc [29] or Men (Figure 2), we evaluated the response of two TNBC cell lines, i.e., MDA-MB-231-shPRDX1 ( Figure 3A) or HCC1806-shPRDX1 ( Figure 3B), or the non-malignant MCF-10A-shPRDX1 ( Figure 3C) cells to the combined Asc/Men treatment as compared to the respective shNTC controls ( Figure 3A-C) or parental cells ( Figure S5). We noticed that both MDA-MB-231-shPRDX1 and HCC1806-shPRDX1 ( Figure 3A,B, respectively) were distinctly more sensitive to incubation with Asc/Men combination as compared to the respective controls ( Figure 3A and Figure S5A for MDA-MB-231 cells and Figure 3B and Figure S5B for HCC1806 cells).…”

Exploring the key communicator role of exosomes in cancer microenvironment through proteomics

“…It is, however, clearly overexpressed in breast cancer tissue relative to normal tissue [70]. Recently, Bajor et al demonstrated that PRDX1 is involved in reducing exogenous oxidative stress and induces cell growth in breast cancer [71]. The H2B1 (Histone H2B type 1-C/3/F/G/I) proteins are related to epigenetic regulation.…”