Journal: Translational Oncology

Article Title: Zeaxanthin targets TOP2A to regulate autophagy and suppress lung cancer progression via the MAPK/ERK pathway

doi: 10.1016/j.tranon.2025.102658

Figure Lengend Snippet: Zea modulates MAPK/ERK signaling to inhibit LC progression. A: Analysis of gene expression differences pre- and post-Zea treatment; B: Heat map analysis of TOP 50 DEGs; C: GO functional enrichment of differentially expressed genes; D: KEGG pathway enrichment of differentially expressed genes; E: WB analysis of MAPK/ERK pathway proteins. A549 cells: NC, Zea, Zea + Ro 67–7476. F: Cell viability measured by CCK-8; G: Cell proliferation assessed by colony formation; H: Cell migration analyzed by Transwell; I: Cell apoptosis detected by flow cytometry; J: WB of autophagy-related proteins; K: IF analysis of autophagosomes and lysosomes, with LC3 for autophagosomes and LAMP1 for lysosomes. ** P < 0.01, *** P < 0.001. The data is presented as mean ± standard deviation, and inter-group comparisons are conducted using ANOVA and Tukey’s post hoc test.

Article Snippet: We also wondered whether the impact of Zea on LC cells was associated with the MAPK/ERK signaling, so we treated A549 cells with Zea and subsequently added MAPK/ERK pathway activator Ro 67–7476 (MCE, USA) to activate this pathway.

Techniques: Gene Expression, Functional Assay, CCK-8 Assay, Migration, Flow Cytometry, Standard Deviation

Journal: Translational Oncology

Article Title: Zeaxanthin targets TOP2A to regulate autophagy and suppress lung cancer progression via the MAPK/ERK pathway

doi: 10.1016/j.tranon.2025.102658

Figure Lengend Snippet: Zea targets TOP2A to enhance autophagy via the MAPK/ERK pathway and impedes LC progression. A549 cells: oe-NC, oe-TOP2A, A: TOP2A mRNA level detected by qRT-PCR A549 cells: oe-NC, oe-TOP2A, oe-NC + Zea, oe-TOP2A + Zea. B: WB analysis of TOP2A, MAPK/ERK pathway proteins (p-ERK1/2, ERK1/2, p-MEK, MEK), and autophagy-related proteins (LC3-I, LC3-II, Beclin 1); C: IF detection of autophagosome and lysosome formation; D: CCK-8 assay of cell viability; E: Colony formation assay of cell proliferation; F: Transwell assay of cell migration; G: Flow cytometry analysis of cell apoptosis. ** P < 0.01, *** P < 0.001. The data is presented as mean ± standard deviation, and inter-group comparisons are conducted using ANOVA and Tukey’s post hoc test.

Article Snippet: We also wondered whether the impact of Zea on LC cells was associated with the MAPK/ERK signaling, so we treated A549 cells with Zea and subsequently added MAPK/ERK pathway activator Ro 67–7476 (MCE, USA) to activate this pathway.

Techniques: Quantitative RT-PCR, CCK-8 Assay, Colony Assay, Transwell Assay, Migration, Flow Cytometry, Standard Deviation

Journal: Translational Oncology

Article Title: Zeaxanthin targets TOP2A to regulate autophagy and suppress lung cancer progression via the MAPK/ERK pathway

doi: 10.1016/j.tranon.2025.102658

Figure Lengend Snippet: In vivo evidence that Zea targets TOP2A to influence the MAPK/ERK pathway and autophagy to mitigate LC progression. Animal groups: oe-NC, oe-TOP2A, oe-NC + Zea, oe-TOP2A + Zea. A: Tumor images from the mouse model; B: Weight of mouse tumor tissues; C: Volume of mouse tumor tissues; D: HE staining images of mouse tumor tissues; E: IHC detection of TOP2A and KI67 expression levels in tumor tissues; F: WB analysis of MAPK/ERK pathway proteins (p-ERK1/2, ERK1/2, p-MEK, MEK) and autophagy-related proteins (LC3-I, LC3-II, Beclin 1); G: IF detection of autophagosome and lysosome colocalization. *** P < 0.001. The data is presented as mean ± standard deviation, and inter-group comparisons are conducted using ANOVA and Tukey’s post hoc test.

Article Snippet: We also wondered whether the impact of Zea on LC cells was associated with the MAPK/ERK signaling, so we treated A549 cells with Zea and subsequently added MAPK/ERK pathway activator Ro 67–7476 (MCE, USA) to activate this pathway.

Techniques: In Vivo, Staining, Expressing, Standard Deviation

Journal: The Journal of Biological Chemistry

Article Title: KRAS G12D mutation promotes pancreatic tumorigenesis by suppressing sirtuin three via the guanine nucleotide exchange factor RCC1

doi: 10.1016/j.jbc.2025.111057

Figure Lengend Snippet: SIRT3 is transcriptionally down-regulated by KRAS G12D . A, differential gene expression of nicotinate and nicotinamide metabolism pathways. B, SIRT3 mRNA levels of HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗∗∗, p < 0.001; ∗∗∗∗, p < 0.0001. C, SIRT3 and KRAS protein levels in HPNE cells with KRAS/Off or KRAS/On for 3 h to 96 h. Cell extracts were analyzed by Western blotting using tubulin as the loading control. Note that the images of KRAS and tubulin bands were derived from the same source images shown in , C and D , as they were from the same experiment. D, effect of SCH772984 on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HPNE cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HPNE KRAS/Off cell extracts were used for comparison. E, effect of SCH772984 on expression of ERK1/2, phosphorylated ERK1/2, and SIRT3 protein levels in KRAS/On HEK293 cells. Cells were treated with SCH772984 at indicated concentrations for 24 h. Cell extracts were analyzed by Western blotting using vinculin as the loading control. HEK293 KRAS/Off cell extracts were used for comparison. F, schematic illustration of the promoter activity assay to identify potential KRAS regulated region in the SIRT3 promoter. G, activity of truncated SIRT3 promoters in HPNE KRAS/Off and HPNE KRAS/On (12 h) cells. The data are presented as mean ± SD ( n = 3). p values were determined by Student’s t test. ∗, p < 0.05;∗∗, p < 0.01. HPNE: human pancreatic normal epithelial cell.

Article Snippet: The ERK phosphorylation inhibitor SCH772984 (HY-50846), MYC inhibitor MCYi361 (HY-129600) and AP-1 inhibitor T-5224 (HY-12270) were purchased from MedChemExpress.

Techniques: Gene Expression, Western Blot, Control, Derivative Assay, Expressing, Comparison, Activity Assay

Journal: bioRxiv

Article Title: An APP-centered molecular gateway integrates innate immunity and retinoic acid signaling to drive irreversible metamorphic commitment

doi: 10.64898/2026.01.22.700939

Figure Lengend Snippet: (A) Settlement rate of inhibitor-treated larvae. The box plots with superimposed jitter plots display the larval settlement rate under various inhibitor treatments. The biofilm stimulus condition is used as the positive control. The concentration of each inhibitor is indicated on the horizontal axis. The data represent the settlement rate of larvae remaining attached out of 10 larvae across six independent biological replicates (total n = 60). Statistical significance among treatment groups was assessed using one-way ANOVA followed by Tukey’s HSD post hoc test, with grouping letters indicating significant differences ( p < 0.05); treatments sharing a letter are not significantly different. (B) Quantitative assessment of the functional hierarchy. The box plots with superimposed jitter plots show the Metamorphic Progression Scores (MPS) for larvae treated with various pharmacological inhibitors with or without all-trans retinoic acid (RA). The MPS was calculated based on the metamorphic stage reached by the larvae in the identical assays used for the settlement rate analysis in (A). The concentration of each inhibitor is indicated on the horizontal axis. The MPS represents the average metamorphic stage reached (0 = brachiolaria; 1 = early; 2 = middle; 3 = late; 4 = pre-juvenile; 5 = juvenile). Statistical significance among the treatment groups was assessed using one-way ANOVA followed by Tukey’s HSD post hoc test ( *p < 0.05; n.s., not significant). A significant RA-dependent rescue condition (a statistically significant increase in MPS compared with the inhibitor-alone condition) is highlighted in grey, establishing the functional hierarchy of the pathways relative to the RA commitment signal. (C) Representative image illustrating pathway functional hierarchy. Images show representative larval morphology under the control, inhibitor-only, and inhibitor + RA conditions. These images specifically represent the high-concentration inhibitor treatments (MyD88 inhibitor: 50 µM; MAPK inhibitors: 10 µM; IKKβ and HSP90AA1 inhibitors: 1 µM). MyD88 inhibition completely blocks the behavioral decision of settlement. JNK and p38 inhibition caused a distinct early-stage arrest (low MPS), and the effects of their inhibition were significantly rescued by RA co-treatment. In contrast, ERK inhibition arrested metamorphosis at the middle stage, and this block was not rescued by exogenous RA. Similarly, IKKβ and HSP90AA1 inhibition arrested metamorphosis at later stages, and this block was not rescued by exogenous RA, functionally placing all three pathways (ERK, IKKβ, and HSP90AA1) downstream of the RA commitment signal. Scale bar: 200 µm. Inhibitors used: T6167923 (MyD88 inhibitor), IKK-16 (IKKβ inhibitor), U0126 (ERK inhibitor), SP600125 (JNK inhibitor), SB202190 (p38 inhibitor), and Luminespib (HSP90AA1 inhibitor).

Article Snippet: The inhibitors were dissolved in DMSO and applied at the indicated concentrations: the MyD88 inhibitor T6167923 (5 or 50 μM; MedChemExpress), the IKKβ inhibitor IKK-16 (0.1 or 1 μM; MedChemExpress), and MAPK inhibitors SP600125 (JNK), SB202190 (p38), and U0126 (ERK) (1 or 10 μM; MedChemExpress or FUJIFILM Wako Pure Chemical Corporation), and the HSP90AA1 inhibitors luminespib (0.1 or 1 μM; Chemscene).

Techniques: Positive Control, Concentration Assay, Functional Assay, Control, Inhibition, Blocking Assay