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