Journal: Acta Pharmaceutica Sinica. B

Article Title: CRISPR-Cas9 gene editing strengthens cuproptosis/chemodynamic/ferroptosis synergistic cancer therapy

doi: 10.1016/j.apsb.2024.05.029

Figure Lengend Snippet: In vitro distribution and tumor-combatting effect of RNP@Cu 2 O@SPF. (A) The viability of cancer and normal cells treated with RNP@Cu 2 O@SPF for 24 h in the dark ( n = 6). (B) CLSM images of SW480 cells incubated with RNP@Cu 2 O@SPF or RNP@Cu 2 O@S (FITC-labelled RNP, RITC-labelled silica). Scale bar, 20 μm. (C) FCM analysis showcasing transfection rates in SW480 cells after 4 h of immersion with PBS, RNP@Cu 2 O@S, or RNP@Cu 2 O@SPF. WB assay (D) and fluorescence imaging (E) of ATP7A expression in SW480 cells treated with RNP@Cu 2 O@S or RNP@Cu 2 O@SPF. Scale bar, 100 μm. (F) Relative copper contents in SW480 cells after a 24 h treatment with Cu 2 O@SPF or RNP@Cu 2 O@SPF ( n = 6). (G) Fluorescence microscopy images of ROS generation in SW480 cells treated with PBS, Cu 2 O@SPF(40 μg/mL), RNP@Cu 2 O@SPF (40 μg/mL), and RNP@Cu 2 O@SPF (80 μg/mL) for 4 h and stained with DCFH-DA to indicate the ROS generation. Scale bar, 50 μm. (H) GSH/GSSG ratio in SW480 cells treated with different concentrations of RNP@Cu 2 O@SPF (25, 50, 100 μg/mL) for 6 h ( n = 6). The data are shown as mean ± SD. ns, no significant; ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: The Cell Copper (Cu) Colorimetric Assay Kit (Elabscience Biotechnology, Wuhan, China) was selected to detect the copper ion content in SW480 cells, and the BCA Protein Colorimetric Assay Kit (Elabscience Biotechnology, Wuhan, China) was also used to determine the total protein concentration.

Techniques: In Vitro, Incubation, Transfection, Fluorescence, Imaging, Expressing, Microscopy, Staining

Journal: Acta Pharmaceutica Sinica. B

Article Title: CRISPR-Cas9 gene editing strengthens cuproptosis/chemodynamic/ferroptosis synergistic cancer therapy

doi: 10.1016/j.apsb.2024.05.029

Figure Lengend Snippet: Anti-tumor mechanisms studies of RNP@Cu 2 O@SPF. (A) The schematic diagram of cell death mechanisms of RNP@Cu 2 O@SPF nanomedicine-mediated synergetic therapy. (B) FCM analysis with Annexin V-FITC/PI dual labels on SW480 cells after 24 h of incubation with different nanoparticles. (C) The percentages of cells undergoing apoptosis (%) in different groups ( n = 3). (D) Viability of SW480 cells grown in media containing either glucose or galactose treated with RNP@Cu 2 O@SPF (ratio 1:1) ( n = 6). (E) Viability of SW480 cells pretreated with 0.1 mmol/L rotenone (Rot), 0.1 mmol/L antimycin A (anti-A), or 1 mmol/L FCCP and then treated with RNP@Cu 2 O@SPF ( n = 6). (F) Viability of SW480 cells post-treated with Antimycin A or UK5099 under different conditions (incubated with various nanoparticles) ( n = 6). (G) Viability of SW480 cells treated with tetrathiomolybdate in different concentrations of RNP@Cu 2 O@SPF (50, 100 μg/mL) ( n = 6). (H) Western blot analysis of DLAT-oligomers, DLAT, and Tubulin expression levels in SW480 cells after treatment with different nanoparticles. (I) Western blot analysis of lipoylated proteins, FDX1, LIAS, ACO-2, SDHB9, and Tubulin expression levels in SW480 cells after treatment with different nanoparticles. (J) Fluorescence depiction of GPX4 levels in SW480 cells post RNP@Cu 2 O@SPF treatment at different dosages (25, 50, and 100 μg/mL) for 24 h. Scale bar, 25 μm. (K) LPO contents in SW480 cells treated with RNP@Cu 2 O@SPF at different concentrations (25, 50, and 100 μg/mL) for 24 h ( n = 6). (L) Survival rates of SW480 cells after 24 h of incubation with solutions of PBS, RNP@Cu 2 O@SPF, PBS+DFO, or RNP@Cu 2 O@SPF+DFO ( n = 6). (M) WB assay of GPX4, ACSL4, and Tubulin expression in SW480 cells treated with RNP@Cu 2 O@SPF at varied concentrations (25, 50, and 100 μg/mL) for 24 h. The data are shown as mean ± SD. ns, not significant; ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: The Cell Copper (Cu) Colorimetric Assay Kit (Elabscience Biotechnology, Wuhan, China) was selected to detect the copper ion content in SW480 cells, and the BCA Protein Colorimetric Assay Kit (Elabscience Biotechnology, Wuhan, China) was also used to determine the total protein concentration.

Techniques: Incubation, Western Blot, Expressing, Fluorescence