Journal: bioRxiv

Article Title: Salvianolic acids are natural senolytics and increase lifespan in old age

doi: 10.64898/2026.04.29.721790

Figure Lengend Snippet: ( a ) A schematic workflow illustrating the preclinical therapeutic procedure. Two weeks after subcutaneous inoculation and in vivo uptake of PC3/PSC27 recombinants, severe combined immunodeficient (M-NSG) mice were subject to either single agent or combinatorial treatment in a metronomic schedule of several cycles. The illustration was created in BioRender. ( b ) Comparative statistics of tumor end volumes. PC3 cancer cells were inoculated either alone or combined with PSC27 stromal cells before being implanted subcutaneously to the hind flank of M-NSG animals, which were administered with MIT and SAA, either alone or in combination. ( c ) Representative images of in vivo senescence in tumor foci by SA-β-gal staining. Scale bar, 50 μm. ( d ) Comparative statistics of tumor senescence as described in ( c ). ( e ) Transcriptional analysis of a subset of SASP factors expressed in epithelial vs. stromal cells acquired from tumor foci after laser capture microdissection (LCM) to isolate stromal and cancer cells, respectively. Signals were normalized to that of the sample with lowest value in the placebo group. ( f ) Transcriptional analysis of SASP factors and two canonical senescence biomarkers p16 INK4a and p21 CIP1 . ( g ) Statistical assessment of DDR and cellular apoptosis in tumors. Values are shown as the percentage of cells positively stained by IF or IHC specific to γH2AX or cleaved caspase 3 (CCL3), respectively. ( h ) Representative images of IHC staining for CCL3 at the completion of treatment regimens. Scale bar, 50 μm. ( i ) Comparative survival of animals sacrificed upon development of advanced bulky disease. Survival duration was calculated from tissue recombinant injection until death. P values were calculated by a two-sided log-rank (Mantel-Cox) test. DDR, DNA damage response. IF, immunofluorescence. IHC, immunohistochemistry. Data in b , d , e , f and g are shown as mean ± SD and representative of 3 independent biological replicates, with P values calculated by Student’s t -tests. ^, P > 0.05; *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: The human prostate epithelial cancer cell line, PC3, was from ATCC and cultured with F-12K media (10% FBS).

Techniques: In Vivo, Staining, Laser Capture Microdissection, Immunohistochemistry, Recombinant, Injection, Immunofluorescence

Journal: The World Journal of Men's Health

Article Title: Inhibition of Ferroptosis in Prostatitis Model by Low Intensity Extracorporeal Shock Wave Therapy through the Integrin-β1/NRF2 Axis

doi: 10.5534/wjmh.250222

Figure Lengend Snippet: Integrin-β1 served as a key mechanoreceptor mediating the anti-ferroptotic action of Li-ESWT. (A–E) Western blot analysis of Integrin-β1, NRF2, and the ferroptosis-related proteins xCT and GPX4 in RWPE-1 cells was performed to evaluate the effect of Li-ESWT-mediated Integrin-β1 activation on NRF2-xCT/GPX4 axis. (F–I) Lipid peroxidation and intracellular iron levels were assessed by flow cytometry using the C11 BODIPY and RhoNOX-6 probes, respectively, to evaluate the occurrence of ferroptosis following Li-ESWT, Integrin-β1 knockdown, or ferroptosis inhibitor Fer-1 treatment. Data were presented as mean±standard deviation. Li-ESWT: low-intensity extracorporeal shock wave therapy, Fer-1: Ferrostatin-1, LPS: lipopolysaccharide. * p<0.05, ** p<0.01, *** p<0.001.

Article Snippet: Human normal prostate epithelial cells RWPE-1 (Procell) were cultured in Prostate Epithelial Cell Medium (ScienCell).

Techniques: Western Blot, Activation Assay, Flow Cytometry, Knockdown, Standard Deviation

Journal: Redox Biology

Article Title: Epithelial redox stress programs macrophage immunometabolism through a ZNF24-MIF–NF–κB pathway in chronic nonbacterial prostatitis

doi: 10.1016/j.redox.2026.104042

Figure Lengend Snippet: Epithelial ROS-ZNF24 axis drives MIF transcription and promotes CD74-dependent M1 macrophage polarization. (A–E) CD74 knockdown attenuates MIF-induced M1 macrophage polarization, as assessed by CD86 and iNOS expression (A–B), proinflammatory cytokine secretion (C), and flow cytometry analysis of F4/80 + CD86 + macrophages (D–E). (F–H) LPS stimulation induces MIF mRNA expression (F), protein expression (G), and secretion (H) in RWPE-1 prostate epithelial cells. (I–K) Transwell co-culture system showing that LPS-stimulated prostate epithelial cells promote M1 polarization and cytokine secretion in iBMDMs, which is suppressed by the MIF inhibitor ISO-1. (L–M) Increased epithelial oxidative stress in EAP mice and LPS-stimulated RWPE-1 cells, indicated by 8-OHdG staining (L) and intracellular ROS levels (M), respectively; NAC effectively reduces ROS accumulation. (N–O) ROS scavenging with NAC suppresses epithelial MIF expression and attenuates M1 marker expression in co-cultured macrophages. (P–T) ZNF24 is induced by epithelial ROS and directly regulates MIF transcription, as shown by ZNF24 expression (P), ZNF24 knockdown (Q), predicted ZNF24 binding motifs in the MIF promoter (R), and ChIP assays demonstrating enhanced ZNF24 binding upon LPS stimulation (S–T). Data are presented as mean ± SD. ns, not significant; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. Abbreviations: iBMDMs, immortalized bone marrow-derived macrophages; RWPE-1, human prostate epithelial cell line; siCD74, small interfering RNA targeting CD74; LPS, lipopolysaccharide; NAC, N-acetylcysteine; ROS, reactive oxygen species.

Article Snippet: The iBMDM cell line (a murine immortalized macrophage line) was kindly provided by academician Feng Shao, and the human prostate epithelial cell line RWPE-1 (Cat. No. CL-0200, Procell) was obtained from Procell. iBMDM cells were cultured in high-glucose DMEM containing 10 % FBS and 1 % penicillin-streptomycin and passaged as needed.

Techniques: Knockdown, Expressing, Flow Cytometry, Co-Culture Assay, Staining, Marker, Cell Culture, Binding Assay, Derivative Assay, Small Interfering RNA

Journal: Current Issues in Molecular Biology

Article Title: Genistein–Butein Co-Treatment Suppresses Glycolytic Metabolism and Induces Apoptotic Signaling in PC-3 Prostate Cancer Cells

doi: 10.3390/cimb48030258

Figure Lengend Snippet: Genistein and butein reduce cell viability in prostate cancer cells. ( A ) Chemical structures of GEN and BTN. ( B ) Viability of PC-3 prostate cancer cells and HPrEC following exposure to increasing concentrations of GEN or BTN for 24 or 48 h, evaluated using the MTT assay. ( C ) Cell viability measured by means of trypan blue exclusion under identical treatment conditions. Results are normalized to untreated controls, and IC 50 values were calculated from 48 h treatment data where indicated. Values represent the mean ± SD of at least three independent experiments; * p < 0.05 versus control. HPrEC: normal human prostate epithelial cells; GEN: genistein; BTN: butein; Conc.: concentration.

Article Snippet: Human prostate cancer PC-3 cells and normal human prostate epithelial cells (HPrEC) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: MTT Assay, Control, Concentration Assay

Journal: Current Issues in Molecular Biology

Article Title: Genistein–Butein Co-Treatment Suppresses Glycolytic Metabolism and Induces Apoptotic Signaling in PC-3 Prostate Cancer Cells

doi: 10.3390/cimb48030258

Figure Lengend Snippet: Combined genistein and butein treatment impairs glycolytic metabolism in PC-3 cells. HPrEC and PC-3 cells were treated with GEN, BTN, or GEN/BTN for 48 h. ( A , B ) Extracellular lactate production and glucose consumption were quantified and normalized to control levels. Enzymatic activities of hexokinase (HK) and pyruvate dehydrogenase (PDH) were assessed following treatment. GEN/BTN co-treatment produced a pronounced suppression of glycolytic activity in PC-3 cells, whereas metabolic parameters in HPrEC cells remained largely unchanged. Data are shown as the mean ± SD (n = 3). Statistical significance is indicated by asterisks (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001). HPrEC: normal human prostate epithelial cells; GEN: genistein; BTN: butein.

Article Snippet: Human prostate cancer PC-3 cells and normal human prostate epithelial cells (HPrEC) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Control, Produced, Activity Assay

Journal: Current Issues in Molecular Biology

Article Title: Genistein–Butein Co-Treatment Suppresses Glycolytic Metabolism and Induces Apoptotic Signaling in PC-3 Prostate Cancer Cells

doi: 10.3390/cimb48030258

Figure Lengend Snippet: Genistein–butein co-treatment decreases ATP levels and clonogenic survival in PC-3 cells. ( A ) Intracellular ATP content was determined after 48 h exposure to GEN, BTN, or GEN/BTN, and expressed relative to control cells (n = 3 independent experiments). ( B ) Crystal violet staining images illustrating treatment-dependent changes in cell survival. ( C ) Quantitative analysis of crystal violet staining measured at 590 nm. Each dot represents an individual replicate, and data are presented as mean ± SD (total n = 8 pooled from at least three independent experiments). Following GEN/BTN co-treatment in PC-3 cells, a significant reduction in ATP production and cell survival was observed, while normal HPrEC cells showed minimal changes. Results represent the mean ± SD from three independent experiments. Statistical significance is indicated by asterisks (* p < 0.05; ** p < 0.01; **** p < 0.0001). HPrEC: normal human prostate epithelial cells; GEN: genistein; BTN: butein.

Article Snippet: Human prostate cancer PC-3 cells and normal human prostate epithelial cells (HPrEC) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Control, Staining

Journal: Current Issues in Molecular Biology

Article Title: Genistein–Butein Co-Treatment Suppresses Glycolytic Metabolism and Induces Apoptotic Signaling in PC-3 Prostate Cancer Cells

doi: 10.3390/cimb48030258

Figure Lengend Snippet: Genistein–butein co-treatment promotes apoptosis and suppresses AKT/ERK signaling in PC-3 cells. ( A ) Apoptotic cell populations were evaluated using Annexin V and viability staining after 48 h treatment. Representative dot plots are shown for HPrEC and PC-3 cells. ( B ) Expression levels of hexokinase II and PDH were analyzed by means of Western blotting, with β-actin serving as a loading control. Corresponding densitometric analyses are presented. ( C ) Phosphorylation status of AKT and ERK, along with levels of cleaved caspase-3 and cleaved PARP, was examined by Western blot analysis. GEN/BTN co-treatment selectively inhibited survival signaling and enhanced apoptotic marker cleavage in PC-3 cells. Data are presented as the mean ± SD. Statistical significance is indicated. Statistical significance is indicated by asterisks (* p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001). HPrEC: normal human prostate epithelial cells; GEN: genistein; BTN: butein; PDH: pyruvate dehydrogenase.

Article Snippet: Human prostate cancer PC-3 cells and normal human prostate epithelial cells (HPrEC) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Staining, Expressing, Western Blot, Control, Phospho-proteomics, Marker