Journal: Acta Biochimica et Biophysica Sinica

Article Title: L-arginine synergistic with 5-fluorouracil intervenes in DNA damage repair via the DNA-PKcs/ATM/ATR pathway in hepatocellular carcinoma cells

doi: 10.3724/abbs.2025137

Figure Lengend Snippet: Co-administration of L-Arg and 5-FU modulates apoptosis and the expression levels of apoptosis-related proteins in HepG2 and HuH-7 cells (A) The apoptotic morphological changes induced by combined L-Arg and 5-FU treatment were evaluated via DAPI staining. (B) The rate of apoptosis resulting from the combination of L-Arg and 5-FU was quantified via Annexin V/FITC-PI staining. (C,D) The effects of L-Arg and 5-FU on the protein expression levels of Bcl-2 and Bax were assessed. (E,F) Immunoblot analysis revealed the effects of L-Arg and 5-FU on the phosphorylation of p53, as well as the cleavage of caspase-9, caspase-3, and PARP. Data are expressed as the mean ± standard deviation, with n = 3. The normal distribution of the data was confirmed prior to analysis. Statistical evaluations were performed via one-way analysis of variance (ANOVA) followed by Dunnett’s t test for pairwise comparisons between groups. Analyses were conducted via GraphPad Prism 10 software. *P < 0.05, **P < 0.01, compared with the control group; # P < 0.05, ## P < 0.01, compared L-Arg with 5-FU group.

Article Snippet: A one-step TUNEL in situ apoptosis detection kit (Red, Elab Fluor® 594), an Annexin V-FITC/PI dual staining kit, and a prestained protein marker (8–180 kDa) were purchased from KeyGEN Biotechnology Company (Nanjing, China).

Techniques: Expressing, Staining, Western Blot, Phospho-proteomics, Standard Deviation, Software, Control

Journal: Acta Biochimica et Biophysica Sinica

Article Title: L-arginine synergistic with 5-fluorouracil intervenes in DNA damage repair via the DNA-PKcs/ATM/ATR pathway in hepatocellular carcinoma cells

doi: 10.3724/abbs.2025137

Figure Lengend Snippet: Co-administration of L-Arg and 5-FU modulates DNA damage and the cell cycle distribution in HepG2 and HuH-7 cells (A) Cell cycle distribution following treatment with L-Arg and 5-FU was analyzed via PI staining. (B) DNA migration after combined treatment with L-Arg and 5-FU was quantified via comet assay. (C) DNA fragmentation induced by the combination of L-Arg and 5-FU was assessed via TUNEL assay. (D) The impact of combined L-Arg and 5-FU treatment on γ-H2AX levels was evaluated via immunofluorescence analysis. (E) Immunoblot analysis further confirmed the effects of L-Arg and 5-FU on γ-H2AX expression levels. Data are expressed as the mean ± standard deviation, with n = 3. The normal distribution of the data was confirmed prior to analysis. Statistical evaluations were performed via one-way analysis of variance (ANOVA) followed by Dunnett’s t test for pairwise comparisons between groups. Analyses were conducted via GraphPad Prism 10 software. *P < 0.05, **P < 0.01, compared with the control group; ## P < 0.01, compared L-Arg with 5-FU group.

Article Snippet: A one-step TUNEL in situ apoptosis detection kit (Red, Elab Fluor® 594), an Annexin V-FITC/PI dual staining kit, and a prestained protein marker (8–180 kDa) were purchased from KeyGEN Biotechnology Company (Nanjing, China).

Techniques: Staining, Migration, Single Cell Gel Electrophoresis, TUNEL Assay, Immunofluorescence, Western Blot, Expressing, Standard Deviation, Software, Control

Journal: Acta Biochimica et Biophysica Sinica

Article Title: L-arginine synergistic with 5-fluorouracil intervenes in DNA damage repair via the DNA-PKcs/ATM/ATR pathway in hepatocellular carcinoma cells

doi: 10.3724/abbs.2025137

Figure Lengend Snippet: The role of reactive oxygen species (ROS) in DNA damage and apoptosis induction during the coadministration of L-Arg and 5-FU (A) Flow cytometric analysis of ROS changes under combined treatment with L-Arg and 5-FU. (B) Fluorescence microscopy-based evaluation of ROS modulation induced by the coadministration of L-Arg and 5-FU. The cells were preincubated with carboxy-PTIO, an NO scavenger (50 μM), for 1 h before subsequent treatments. (C) Effects of the ROS inhibitor N-acetylcysteine (NAC) on the protein expression levels of p-AKT, DNA-PKcs, p-ATM, p-ATR, γ-H2AX, and cleaved caspase-3. The cells were pre-treated with NAC (3 mM) for 1 h prior to subsequent treatments. Data are expressed as the mean ± standard deviation, with n = 3. The normal distribution of the data was confirmed prior to analysis. Statistical evaluations were performed via one-way analysis of variance (ANOVA) followed by Dunnett’s t test for pairwise comparisons between groups. Analyses were conducted via GraphPad Prism 10 software. *P < 0.05, **P < 0.01, compared with the control group; # P < 0.05, ## P < 0.01, compared L-Arg with 5-FU group.

Article Snippet: A one-step TUNEL in situ apoptosis detection kit (Red, Elab Fluor® 594), an Annexin V-FITC/PI dual staining kit, and a prestained protein marker (8–180 kDa) were purchased from KeyGEN Biotechnology Company (Nanjing, China).

Techniques: Fluorescence, Microscopy, Expressing, Standard Deviation, Software, Control

Journal: Acta Biochimica et Biophysica Sinica

Article Title: L-arginine synergistic with 5-fluorouracil intervenes in DNA damage repair via the DNA-PKcs/ATM/ATR pathway in hepatocellular carcinoma cells

doi: 10.3724/abbs.2025137

Figure Lengend Snippet: iNOS affects the levels of apoptosis-related and DNA damage repair-related proteins in HepG2 and HuH-7 cells (A) HepG2 cells. (B) HuH-7 cells. Data are expressed as the mean ± standard deviation, with n = 3. The normal distribution of the data was verified prior to analysis. Statistical evaluations were performed via one-way analysis of variance (ANOVA), and Dunnett’s t test was used for pairwise comparisons between groups. Analyses were conducted via GraphPad Prism 10 software. **P < 0.01, compared with the control group; # P < 0.05, ## P < 0.01, compared L-Arg with 5-FU group.

Article Snippet: A one-step TUNEL in situ apoptosis detection kit (Red, Elab Fluor® 594), an Annexin V-FITC/PI dual staining kit, and a prestained protein marker (8–180 kDa) were purchased from KeyGEN Biotechnology Company (Nanjing, China).

Techniques: Standard Deviation, Software, Control

Journal: Acta Biochimica et Biophysica Sinica

Article Title: L-arginine synergistic with 5-fluorouracil intervenes in DNA damage repair via the DNA-PKcs/ATM/ATR pathway in hepatocellular carcinoma cells

doi: 10.3724/abbs.2025137

Figure Lengend Snippet: Co-administration of L-Arg and 5-FU modulates the expression levels of proteins associated with DNA damage repair, the PI3K/AKT signaling pathway, and apoptosis in rat liver tissue (A) Immunofluorescence staining was used to analyze the effect of combined L-Arg and 5-FU treatment on DNA-PKcs levels. (B) Immunoblot analysis was used to evaluate effects of co-administration of L-Arg and 5-FU on the levels of DNA-PKcs, phosphorylated ATM (p-ATM), phosphorylated ATR (p-ATR), and phosphorylated AKT (p-AKT) in rat liver tissue. (C) The influence of coadministration of L-Arg and 5-FU on the levels of phosphorylated p53 (p-p53), Bcl-2, cleaved caspase-3, and cleaved caspase-9 in rat liver tissue was assessed via immunoblot analysis. Data are expressed as the mean ± standard deviation, with n = 12. The normal distribution of the data was confirmed prior to analysis. Statistical evaluations were performed via one-way analysis of variance (ANOVA) followed by Dunnett’s t test for pairwise comparisons between groups. Analyses were conducted via GraphPad Prism 10 software.**P < 0.01, compared with the normal control group; ΔΔ P<0.01, compared with the DEN model group. ## P < 0.01, compared L-Arg with 5-FU group.

Article Snippet: A one-step TUNEL in situ apoptosis detection kit (Red, Elab Fluor® 594), an Annexin V-FITC/PI dual staining kit, and a prestained protein marker (8–180 kDa) were purchased from KeyGEN Biotechnology Company (Nanjing, China).

Techniques: Expressing, Immunofluorescence, Staining, Western Blot, Standard Deviation, Software, Control