Journal: Cell Death Discovery

Article Title: AURKA/PHB2 signaling drives acquired resistance to KRAS G12C inhibitors in KRAS G12C -mutant NSCLC

doi: 10.1038/s41420-026-03080-4

Figure Lengend Snippet: A Intersection of prognostic risk genes, tumor-normal DEGs, and Sotorasib resistance genes revealed AURKA as the sole overlapping gene. B Boxplot showing AURKA expression levels in Sotorasib-resistant versus sensitive tumors from 16 Kras-G12C/Trp53 genetically engineered mice (** p < 0.01, Wilcoxon test). C qPCR detection of AURKA expression in normal lung epithelial cells and KRAS G12C mutant lung cancer cells H358 and Calu-1. D – G MTS assay for the proliferation of Sotorasib-sensitive H358 cells and Sotorasib-resistant H358-SR at various concentrations of Sotorasib, and calculation of IC50 by SPSS27. H Kaplan-Meier survival curves for TCGA-LUAD patients stratified by AURKA expression (high vs. low; log-rank p < 0.001). I Immunohistochemical (IHC) expression of AURKA in tumor tissues versus normal tissues from lung adenocarcinoma patients with KRAS G12C mutation. J, K qPCR and WB for the assessment of the expression levels of AURKA and PHB2 in H358, Calu-1, H358-SR and Calu-1-SR cells. * p < 0.05; **** p < 0.0001.

Article Snippet: Sections were incubated overnight at 4 °C with primary antibodies against AURKA (1:100; MCE, Cat. No. HY- P80971 ), PHB2 (1:100; MCE, Cat. No. HY- P82103 ), and Ki-67 (1:500; CST, Cat. No. #9449), followed by incubation with a secondary antibody and horseradish peroxidase (HRP)-conjugated streptavidin for 15 min. Immunoreactivity was visualized using DAB, and nuclei were counterstained with hematoxylin.

Techniques: Expressing, Mutagenesis, MTS Assay, Immunohistochemical staining

Journal: Cell Death Discovery

Article Title: AURKA/PHB2 signaling drives acquired resistance to KRAS G12C inhibitors in KRAS G12C -mutant NSCLC

doi: 10.1038/s41420-026-03080-4

Figure Lengend Snippet: A Whole Calu-1 lysates were generated for immunoprecipitation with anti-AURKA antibodies or a control IgG, followed by LC-MS/MS analysis of the immunocomplexes. A total of 72 potential interacting proteins with high fidelity were identified. PHB2 emerged as a possible AURKA-interacting protein. B Endogenous co-immunoprecipitation assays were conducted to explore the interaction between AURKA and PHB2 in Calu - 1 cells. C Representative immunofluorescence images were obtained to demonstrate the colocalization of AURKA and PHB2 in H358 and Calu - 1 cells. D , E qRT-PCR was employed to detect the mRNA expression levels of PHB2 or AURKA in Calu-1 or Calu-1-SR cells. These cells had either stable knockdown or overexpression of AURKA or PHB2. F WB was utilized to detect the protein expression levels of PHB2 or AURKA in Calu - 1 or Calu - 1 - SR cells with stable knockdown or overexpression of AURKA or PHB2. G , H A luciferase promoter assay was performed.

Article Snippet: Sections were incubated overnight at 4 °C with primary antibodies against AURKA (1:100; MCE, Cat. No. HY- P80971 ), PHB2 (1:100; MCE, Cat. No. HY- P82103 ), and Ki-67 (1:500; CST, Cat. No. #9449), followed by incubation with a secondary antibody and horseradish peroxidase (HRP)-conjugated streptavidin for 15 min. Immunoreactivity was visualized using DAB, and nuclei were counterstained with hematoxylin.

Techniques: Generated, Immunoprecipitation, Control, Liquid Chromatography with Mass Spectroscopy, Immunofluorescence, Quantitative RT-PCR, Expressing, Knockdown, Over Expression, Luciferase, Promoter Assay

Journal: Cell Death Discovery

Article Title: AURKA/PHB2 signaling drives acquired resistance to KRAS G12C inhibitors in KRAS G12C -mutant NSCLC

doi: 10.1038/s41420-026-03080-4

Figure Lengend Snippet: A Immunoblot analysis of PHB2, AURKA, and β-actin in CHX-treated (10 µg/mL) Calu-1 cells stably expressing AURKA at specified intervals. The half - life of PHB2 was evaluated. B, C WB was used to detect the expression of PHB2 and AURKA in Calu - 1 cells. These cells had stable knockdown of AURKA or were treated with siPHB2. They were then treated with CHX and MG132 (a proteasome inhibitor, 10 μM) for 8 h. D GSEA enrichment plot of the PI3K-Akt signaling pathway in AURKA-knockdown tumor samples compared to control tumors. The pathway was significantly suppressed (NES = −1.58, p < 0.001, adjusted p < 0.05), with a distinct depletion of pathway genes at the lower end of the ranked DEG list. E Gene ratio plot illustrating the proportion of pathway genes in the PI3K-Akt signaling pathway relative to other enriched pathways (adjusted p < 0.05). F Over-Representation Analysis (ORA) of KEGG pathways showing significant suppression of oncogenic pathways and activation of immune-related pathways following AURKA knockdown ((adjusted p < 0.05). G Detection of the expression of AKT and p - AKT in Calu - SR cells of the NC, AURKA - SH and AURKA - SH/PHB2 - OE groups after 24-h treatment with Sotorasib by WB.

Article Snippet: Sections were incubated overnight at 4 °C with primary antibodies against AURKA (1:100; MCE, Cat. No. HY- P80971 ), PHB2 (1:100; MCE, Cat. No. HY- P82103 ), and Ki-67 (1:500; CST, Cat. No. #9449), followed by incubation with a secondary antibody and horseradish peroxidase (HRP)-conjugated streptavidin for 15 min. Immunoreactivity was visualized using DAB, and nuclei were counterstained with hematoxylin.

Techniques: Western Blot, Stable Transfection, Expressing, Knockdown, Control, Activation Assay

Journal: Cell Death Discovery

Article Title: AURKA/PHB2 signaling drives acquired resistance to KRAS G12C inhibitors in KRAS G12C -mutant NSCLC

doi: 10.1038/s41420-026-03080-4

Figure Lengend Snippet: A , B qPCR and WB were employed to measure the expression level of PHB2 in Calu-1-SR/NC, Calu-1-SR/AURKA-SH and Calu-1-SR/AURKA-SH/PHB2-OE cells. C MTS was used to detect the cell viability of Calu-1-SR/NC, Calu-1-SR/AURKA-SH and Calu-1-SR/AURKA-SH/PHB2-OE cells after treatment with Sotorasib, and IC50 was calculated by SPSS27. D , E Colony formation assay was used to detect the cell proliferation of Calu-1-SR/NC, Calu-1-SR/AURKA-SH and Calu-1-SR/AURKA-SH/PHB2-OE cells after treatment with Sotorasib. F Assay for Caspase 3/7 activity was employed to measure the apoptosis of Calu-1-SR/NC, Calu-1-SR/AURKA-SH and Calu-1-SR/AURKA-SH/PHB2-OE cells treated with Sotorasib. G Western blot was used to detect the expression of apoptosis-related proteins Bcl-2, Bax and Bim in Calu-1-SR/NC, Calu-1-SR/AURKA-SH and Calu-1-SR/AURKA-SH/PHB2-OE cells treated with Sotorasib. H Transwell assay was used to determine the migration ability of Calu-1-SR/NC, Calu-1-SR/AURKA-SH and Calu-1-SR/AURKA-SH/PHB2-OE cells. I Schematic diagram illustrating AURKA/PHB2-mediated regulation of KRAS G12C resistance to Sotorasib.

Article Snippet: Sections were incubated overnight at 4 °C with primary antibodies against AURKA (1:100; MCE, Cat. No. HY- P80971 ), PHB2 (1:100; MCE, Cat. No. HY- P82103 ), and Ki-67 (1:500; CST, Cat. No. #9449), followed by incubation with a secondary antibody and horseradish peroxidase (HRP)-conjugated streptavidin for 15 min. Immunoreactivity was visualized using DAB, and nuclei were counterstained with hematoxylin.

Techniques: Expressing, Colony Assay, Activity Assay, Western Blot, Transwell Assay, Migration

Journal: Cell Death Discovery

Article Title: AURKA/PHB2 signaling drives acquired resistance to KRAS G12C inhibitors in KRAS G12C -mutant NSCLC

doi: 10.1038/s41420-026-03080-4

Figure Lengend Snippet: A , D H358-SR or Calu-1-SR cells underwent treatment with the specified concentrations of the AURKA inhibitor Alisertib for 48 h, and the expression level of AURKA measured via Western Blot analysis. B , E Calu-1-SR cells were treated with Sotorasib and Alisertib for 48 h; cell viability and inhibition rate were assessed using MTS. C , F The score of the drug synergy experiment was determined by the Bliss model and displayed as a heatmap. G , I 5 × 10 6 Calu-1-SR cells were injected subcutaneously into nude mice. After allowing palpable tumors to develop for 7 days, the mice were randomly assigned to four groups. These groups were treated with vehicle control (0.01% DMSO in PBS), Sotorasib (30 mg/kg/d), Alisertib (30 mg/kg/d), or a combination of Sotorasib and Alisertib administered every other day ( n = 6 per group). G The tumor size was measured at specified time intervals and calculated. The tumor volume was computed using the formula: V = 1/2 × larger diameter × (smaller diameter) 2 . Growth curves were then plotted using the average tumor volume within each experimental group at the set time points. H At the conclusion of the treatment, the tumors were excised. I The weights of the tumors were measured. J Sotorasib and/or Alisertib administration does not influence the weight of the mice. K Calu-1-SR xenograft tissues obtained from mice were fixed, sliced into sections, and then mounted on glass slides. Tumor samples were processed for IHC staining using Abs against Ki-67, AURKA, and PHB2.

Article Snippet: Sections were incubated overnight at 4 °C with primary antibodies against AURKA (1:100; MCE, Cat. No. HY- P80971 ), PHB2 (1:100; MCE, Cat. No. HY- P82103 ), and Ki-67 (1:500; CST, Cat. No. #9449), followed by incubation with a secondary antibody and horseradish peroxidase (HRP)-conjugated streptavidin for 15 min. Immunoreactivity was visualized using DAB, and nuclei were counterstained with hematoxylin.

Techniques: Expressing, Western Blot, Inhibition, Injection, Control, Immunohistochemistry

Journal: iScience

Article Title: GUTK induces apoptosis in reactivating quiescent prostate cancer cells via Aurora A-mediated stabilization of SOD2

doi: 10.1016/j.isci.2026.115739

Figure Lengend Snippet: GUTK downregulates Aurora A expression during the reactivation of quiescent PCa cells (A and B) RT-PCR analysis of SOD2 mRNA levels in quiescent LNCaP (A) and DU145 (B) cells treated with or without 20 μM GUTK for the indicated times during cell cycle re-entry. (C and D) Quiescent LNCaP and DU145 cells were induced to re-enter the cell cycle and simultaneously exposed to 10 μM MG132 or DMSO for the indicated times. Immunoblot analysis of SOD2 protein was performed in quiescent LNCaP (C) and DU145 (D) cells. GAPDH served as the loading control. Quantification of relative protein levels is shown in the lower panel. (E and F) Quiescent LNCaP and DU145 cells were induced to re-enter the cell cycle for 6 h and then exposed to 50 μM CHX or DMSO for the indicated times. Immunoblot analysis of SOD2 and Aurora A proteins was performed in quiescent LNCaP (E) and DU145 (F) cells. β-actin or GAPDH served as loading control. Quantification of relative protein levels is shown in the lower panel. (G and H) Immunoblot analysis of Aurora A protein levels in quiescent LNCaP (G) and DU145 (H) cells treated with 20 μM GUTK for the indicated times during cell cycle re-entry. β-actin served as loading control. Quantification of relative protein levels is shown in the lower panel. (I and J) RT-PCR analysis of Aurora A mRNA levels in quiescent LNCaP (I) and DU145 (J) cells treated with 20 μM GUTK for the indicated times during cell cycle re-entry. Cont: proliferating control cells; Qsct: quiescent cells; CHX: cycloheximide. Data are presented as mean ± SD from three independent experiments. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus indicated group. ns: not significant.

Article Snippet: Sections were subjected to hematoxylin and eosin (H&E) staining or immunohistochemical staining using primary antibodies against Ki-67 (ab16667, Abcam, Shanghai, China), SOD2 (ET1701-54, HUABIO, Hangzhou, China), Cleaved Caspase 3 (#9661, Cell Signaling Technology, Danvers, USA) and Aurora A (ET1609-22, HUABIO, Hangzhou, China).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Control

Journal: iScience

Article Title: GUTK induces apoptosis in reactivating quiescent prostate cancer cells via Aurora A-mediated stabilization of SOD2

doi: 10.1016/j.isci.2026.115739

Figure Lengend Snippet: Aurora A overexpression suppresses SOD2 expression and attenuates GUTK-induced apoptosis in reactivating quiescent PCa cells (A and B) RT-PCR validation of Aurora A mRNA overexpression in LNCaP (A) and DU145 (B) cells stably transfected with either empty vector (EV) or an Aurora A expression construct (Aurora A OE). (C and D) Immunoblot analysis of Aurora A and SOD2 protein levels in quiescent EV control or Aurora A OE LNCaP (C) and DU145 (D) cells at the indicated times after cell cycle re-entry. GAPDH served as the loading control. Quantification of relative protein levels is shown in the right panel. (E and F) Apoptosis analysis by Annexin V-FITC/PI staining and flow cytometry in EV control or Aurora A OE LNCaP (E) and DU145 (F) cells treated with or without 20 μM GUTK for the indicated times during cell cycle re-entry. Quantification of apoptotic cell percentages is shown in the right panel. (G and H) ΔΨm analysis by JC-1 staining and flow cytometry in quiescent EV control and Aurora A OE LNCaP (G) and DU145 (H) cells treated with or without 20 μM GUTK for the indicated times during cell cycle re-entry. Quantification of cells with low ΔΨm is shown in the right panel. ΔΨm: mitochondrial membrane potential. Data are presented as mean ± SD from three independent experiments. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 versus indicated group.

Article Snippet: Sections were subjected to hematoxylin and eosin (H&E) staining or immunohistochemical staining using primary antibodies against Ki-67 (ab16667, Abcam, Shanghai, China), SOD2 (ET1701-54, HUABIO, Hangzhou, China), Cleaved Caspase 3 (#9661, Cell Signaling Technology, Danvers, USA) and Aurora A (ET1609-22, HUABIO, Hangzhou, China).

Techniques: Over Expression, Expressing, Reverse Transcription Polymerase Chain Reaction, Biomarker Discovery, Stable Transfection, Transfection, Plasmid Preparation, Construct, Western Blot, Control, Staining, Flow Cytometry, Membrane

Journal: iScience

Article Title: GUTK induces apoptosis in reactivating quiescent prostate cancer cells via Aurora A-mediated stabilization of SOD2

doi: 10.1016/j.isci.2026.115739

Figure Lengend Snippet: GUTK combined with docetaxel enhances the reduction of orthotopic prostate tumor growth in C57BL/6 mice (A) Experimental schematic of the orthotopic prostate cancer model and treatment schedule in C57BL/6 mice established using RM-1 murine PCa cells. Mice were randomized into four groups: vehicle, GUTK, docetaxel, or combination treatment ( n = 6 per group). (B) Representative images of major organ morphology (heart, liver, spleen, lungs, kidneys) collected on day 9. (C) Daily body weight measurements throughout the experiment period. (D) Representative images of dissected tumors collected at the day 9 endpoint. (E) Tumor volume quantification at the endpoint. (F) Representative immunohistochemical staining of tumor sections for H&E, Ki-67, SOD2, Aurora A and cleaved caspase 3. Scale bars, 50 μm. Quantification of relative protein levels is shown in the lower panel. Solvent A represents Tween 80: ethanol: saline = 20: 13: 67. Solvent B represents Cremophor EL: Ethanol: 5% glucose = 1: 1: 38. Data are presented as mean ± SEM for each group. ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus vehicle control group.

Article Snippet: Sections were subjected to hematoxylin and eosin (H&E) staining or immunohistochemical staining using primary antibodies against Ki-67 (ab16667, Abcam, Shanghai, China), SOD2 (ET1701-54, HUABIO, Hangzhou, China), Cleaved Caspase 3 (#9661, Cell Signaling Technology, Danvers, USA) and Aurora A (ET1609-22, HUABIO, Hangzhou, China).

Techniques: Immunohistochemical staining, Staining, Solvent, Saline, Control

Journal: Cell Stress & Chaperones

Article Title: Proteomic and phenotypic profiling of replicative-senescent human foreskin fibroblasts under brief heat shock

doi: 10.1016/j.cstres.2026.100174

Figure Lengend Snippet: Expression of several senescence-associated proteins under HS conditions at 41 °C and 45 °C for 30 min in aged HFF-1 cells. (a) Detection of protein expression levels of GORAB, AURKB, RhoC, α-N-catenin, RRM2, AURKA, and PLK1 under 41 °C and 45 °C HS. Protein expression was quantified as fold-change relative to untreated controls (set to 1), normalized to GAPDH. (b) GORAB. (c) AURKB. (d) RhoC. (e) α-N-catenin. (f) RRM2. (g) AURKA. (h) PLK1. Representative blots were cropped only to improve presentation; the corresponding full-length, uncropped blots with molecular-weight markers and the associated GAPDH loading control are provided in Supplementary Figure S1 (the first lane corresponds to an unrelated sample and was not analyzed). Data are representative of three independent experiments, with results presented as mean ± SD. ** P < 0.01, *** P < 0.001, **** P < 0.0001, compared with the control group.

Article Snippet: Membranes were blocked with 5% nonfat milk for 1 h at room temperature and incubated overnight at 4 °C with primary antibodies against GORAB (Proteintech, Wuhan, China; 17798–1-AP; 1:1000), AURKA (Cell Signaling Technology, Danvers, MA, USA; 14475T; 1:1000), AURKB (Cell Signaling Technology; 28711T; 1:1000), RhoC (Cell Signaling Technology; 3430T; 1:1000), RRM2 (Cell Signaling Technology; 65939T; 1:2000), PLK1 (Cell Signaling Technology; 4513T; 1:1000), α-N-catenin (Cell Signaling Technology; 2163T; 1:1000), and GAPDH (Abcam, Cambridge, UK; ab125247; 1:10,000) as the loading control.

Techniques: Expressing, Molecular Weight, Control