Journal: Cell death & disease

Article Title: POLQ inhibition attenuates the stemness and ferroptosis resistance in gastric cancer cells via downregulation of dihydroorotate dehydrogenase.

doi: 10.1038/s41419-024-06618-5

Figure Lengend Snippet: Fig. 2 POLQ regulates the cancer stem cell-like characteristics of GC cells through DHODH. A Heatmap showing differently expressed genes in control and POLQ-knockdown GC cells. Values are scaled as indicated (2 to −2; n = 3). B Heatmap showing differently expressed genes in POLQlow versus POLQhigh stomach adenocarcinoma. C Public database [https://depmap.org/portal/] correlation analysis in cell lines (n = 84) between POLQ and DHODH. Significance was evaluated by Pearson correlation. D, E The mRNA and protein levels of DHODH were detected in control and POLQ- knockdown cell lines. F, G The mRNA and protein levels of DHODH were detected in GC cells treated with vehicle or NVB (100 µM) for 24 h. H Protein expression levels in POLQ- knockdown cells overexpressing DHODH. I Sphere formation in control (ShNC), POLQ knockdown (ShPOLQ), and ShPOLQ + DHODH cells was analyzed using oncosphere-initiating medium. J The serial sphere- forming capacity. K The mRNA levels of gastric cancer stem cell markers in control (ShNC), POLQ knockdown (ShPOLQ), and ShPOLQ + DHODH cells. Results are shown as mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: 30 μg of protein was used for immunoblot analysis using antibodies against DHODH (1:1000; Proteintech), POLQ (1:1000; Affinity), 4HNE (1 μg/ ml; Jaica), and β-actin (1:1000; Proteintech).

Techniques: Control, Knockdown, Expressing

Journal: Cell death & disease

Article Title: POLQ inhibition attenuates the stemness and ferroptosis resistance in gastric cancer cells via downregulation of dihydroorotate dehydrogenase.

doi: 10.1038/s41419-024-06618-5

Figure Lengend Snippet: Fig. 4 Upregulation the expression of DHODH blocks ferroptosis susceptibility mediated by POLQ inhibition. A Cell viability in control (ShNC), POLQ knockdown (ShPOLQ), and ShPOLQ + DHODH cells treated with RSL3 (10 µM) or erastin (15 µM) for 24 h. B-E Respective lipid peroxidation levels were assessed and calculated by flow cytometry using BODIPY C11. F The mRNA levels of PTGS2 were detected in control (ShNC), POLQ knockdown (ShPOLQ), and ShPOLQ + DHODH GC cells treated with RSL3 (5 µM) or erastin (10 µM) for 24 h. Results are shown as mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: 30 μg of protein was used for immunoblot analysis using antibodies against DHODH (1:1000; Proteintech), POLQ (1:1000; Affinity), 4HNE (1 μg/ ml; Jaica), and β-actin (1:1000; Proteintech).

Techniques: Expressing, Inhibition, Control, Knockdown, Cytometry

Journal: Cell death & disease

Article Title: POLQ inhibition attenuates the stemness and ferroptosis resistance in gastric cancer cells via downregulation of dihydroorotate dehydrogenase.

doi: 10.1038/s41419-024-06618-5

Figure Lengend Snippet: Fig. 7 Sulfasalazine with POLQ inhibition efficiently facilitates treatment in gastric cancer. A Schematic of subcutaneous xenograft model treated with vehicle, novobiocin, sulfasalazine, or sulfasalazine in combination with novobiocin (n = 4). B Morphological images of the tumors. C The weights of subcutaneous tumors in indicated mice. D Mice weights were recorded after vehicle, novobiocin, sulfasalazine, or sulfasalazine in combination with novobiocin treatment. E, F IHC staining of DHODH, 4HNE, and MKi67 in subcutaneous tumors derived from indicated mice (scale bar = 50 μm). The expression of PTGS2 (G), CD24, and CD44 (H) in subcutaneous tumors from indicated mice. I, J BLI and images of intraperitoneal tumor formation model treated with vehicle, novobiocin, sulfasalazine, or sulfasalazine in combination with novobiocin (n = 4). Results are shown as mean ± SD. *P < 0.05; **P <0.01; ***P < 0.001; ns not significant.

Article Snippet: 30 μg of protein was used for immunoblot analysis using antibodies against DHODH (1:1000; Proteintech), POLQ (1:1000; Affinity), 4HNE (1 μg/ ml; Jaica), and β-actin (1:1000; Proteintech).

Techniques: Inhibition, Immunohistochemistry, Derivative Assay, Expressing