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: Journal of Experimental & Clinical Cancer Research : CR

Article Title: 3-O-acetyl-11-keto-β-boswellic acid exerts anti-tumor effects in glioblastoma by arresting cell cycle at G2/M phase

doi: 10.1186/s13046-018-0805-4

Figure Lengend Snippet: Proposed mechanistic scheme: AKBA suppresses glioblastoma by arresting cell cycle at G2/M phase. AKBA activates p21 which decreases expression of FOXM1 and its downstream target genes including Aurora A, Aurora B, PLK1, and cyclin B1. Inhibiting Aurora A suppresses the expression of PLK1 which leads to the inactivation of CDK1/cyclin B1 complex and arrests the cell cycle at the G2/M checkpoint. Inhibiting Aurora B decreases expression of TOP2A and inhibits mitosis

Article Snippet: After blocking with 5% fat-free milk in TBST for 1 h, membranes were immunoblotted with primary antibodies to caspase3, cleaved-caspase3, PARP, cleaved-PARP, p21, p-RB, PLK1, p-CDK1, and Aurora B (Cell Signaling Technology); p53, FOXM1, Aurora A, CDC-25C, CDK-1, cyclin B1, TOP2A, and GAPDH (Proteintech) at the appropriate dilutions with gentle shaking overnight at 4 °C.

Techniques: Expressing