Journal: Biochemistry and Biophysics Reports
Article Title: Anti-tumor analysis of the RIG-I agonist in vitro and in vivo
doi: 10.1016/j.bbrep.2025.102249
Figure Lengend Snippet: nCoV-L induced cancer cell death in vitro and suppressed tumor growth in viv o (A) Schematic structure of the stem-loop-structured nCoV-L RNA agonist. (B, C) nCoV-L activated the RIG-I signalling in cancer cells. Hep3B (B) or Huh7 (C) cells were collected and subjected to Western blot analysis after treated with Lipo or Lipo mixed with 2 μg nCoV-L for 24 h. Protein levels of RIG-I, p-IRF3, STAT1, p-STAT1, p–NF–κb and ISG56 were compared to control and Lipo groups. (D) RIG-I signalling deficiency suppressed nCoV-L-induced cell death. Huh7.5.1 cells were transfected with 50 ng nCoV-L. Cell viability was measured 24 h post-transfection using a CCK-8 assay. Data represented mean ± SD; P < 0.05. (E) Ablation of 5ʹ-PPP abolishes nCoV-L-induced cell death. Hep3B cells were transfected with 200 ng of in vitro -transcribed nCoV-L RNA or 5ʹ-PPP-deficient synthetic nCoV-L RNA. Cell viability was quantified 24 h later by CCK-8 assay. Data represented mean ± SD; P < 0.0001. (F) Dose-dependent cytotoxicity of nCoV-L across carcinoma cell lines. Hep3B, NCI–H460, SW620, SW480, and CT26.WT cells were transfected with 50, 100, or 150 ng nCoV-L. Cell viability was assessed 48 h post-transfection using CCK-8 assay. (G) nCoV-L suppressed NCI–H1299 xenograft growth. Mice received intratumoral injections of PBS, lipo, or 10 μg nCoV-L complexed with lipo (nCoV-L/lipo) for 5 consecutive days (arrows, days 0–4). Tumor volumes are shown as mean ± SEM (n = 5); P < 0.0001 vs. PBS (two-way ANOVA). (H) Representative excised tumors from (G). Scale bar: 0.5 cm.
Article Snippet: Antibodies against cleaved-PARP (cat#5625S), cleaved-Caspase3 (cat#9661S), PUMA (cat#98672), p-IRF3 (cat#29047), STAT1 (cat#14994), p-STAT1 (cat#9167), p–NF–κb (cat#3033), ISG56 (cat#14769) and β-actin (cat#4967) were purchased from Cell Signaling Technology.
Techniques: In Vitro, Western Blot, Control, Transfection, CCK-8 Assay
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