mouse prostate primary epithelial cells  (ATCC)

Journal: Frontiers in Immunology

Article Title: Hsp90β inhibition upregulates interferon response and enhances immune checkpoint blockade therapy in murine tumors

doi: 10.3389/fimmu.2022.1005045

Figure Lengend Snippet: Hsp90β-selective inhibitor NDNB1182. (A) Association of high HSP90AB1 expression with worse PSA progression free survival (PFS) and overall survival, with data extracted from the database of a phase 2 clinical trial of ipilimumab in metastatic castration-resistant prostate cancer *P<0.05, log rank test between the two cohorts . (B) Association of high HSP90AB1 expression with worse overall survival for anti-PD1 or anti-PD-L1 treatment with data from multiple clinical studies, drawn with KMplot. (C) The generic structure of NDNB1182 and IC50 values against Hsp90 isoforms determined using fluorescence polarization (FP) assay. (D) Western blot of Src in mouse prostate cancer cell line PPS treated with DMSO, NDNB1182 or luminespib for 12 hours, with the semi-quantitative result presented on the right. (E) HPLC result confirming the high purity of NDNB1182. (F) Dose-response curves and IC50 values of NDNB1182 on four cancer cell lines. Nonlinear regression modeling with log(inhibitor) and normalized response of variable slopes was conducted in Graphpad Prism. (G) Dose-response curves and IC50 values of NDNB1182 on primary prostate epithelial cells grown from wild type C57BL/6 mice and the mouse fibroblast cell line L cells. (H) HSP90AB1 transcript levels for prostate-lineage cell lines in the Depmap database portal with the plot generated by Depmap.

Article Snippet: As comparison, the effect of NDNB1182 was tested on two normal cell types, mouse prostate primary epithelial cells (grown from the dissociated prostate glands of wild type C57BL/6 mice) and the mouse fibroblast cell line L cells (ATCC, CRL-2648).

Techniques: Expressing, Fluorescence, FP Assay, Western Blot, Generated

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-derived MIF is upregulated in CNP and is correlated with inflammation severity. (A) ELISA quantification of MIF protein levels in plasma samples from CP-LS patients (n = 40) and healthy controls (CTR, n = 13). (B) Correlation between MIF concentration and disease duration in CP-LS patients (Spearman's r = 0.4770, p < 0.001). (C) Representative H&E staining and MIF IHC of prostate tissues from BPH patients with mild, moderate, and severe inflammation, showing increasing MIF expression with inflammatory severity. (D) t-SNE plot of single-cell transcriptomic data from normal and BPH human prostate tissues, showing clustering of major cell types. (E) Violin plots of MIF expression across cell types, showing enrichment in epithelial cells. (F) t-SNE feature plots showing overlapping expression of MIF and the epithelial marker EPCAM, with enrichment in epithelial cells. (G) Immunofluorescence staining of BPH tissues showing co-localization of MIF (green) and EPCAM (red) with increasing inflammatory severity. (H) ELISA quantification of MIF levels in serum samples from EAP and CTR mice (n = 5/group). (I) Behavioral assessment of pelvic pain by von Frey testing in EAP versus CTR mice, showing increased mechanical allodynia in EAP. (J) Representative H&E staining of mouse prostate tissues showing inflammatory infiltrates in EAP. (K) Quantification of prostate inflammation scores in CTR and EAP mice (n = 5/group). (L) IHC staining of mouse prostate tissues showing elevated MIF expression in EAP. Data are presented as the mean ± SD. ns, not significant; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. Abbreviations: CP-LS, chronic prostatitis-like symptoms; CTR, control; EAP, experimental autoimmune prostatitis; t-SNE, t-distributed stochastic neighbor embedding.

Article Snippet: Mouse primary prostate epithelial cells were obtained from Procell (Cat. No. CP-M064, Procell).

Techniques: Derivative Assay, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Concentration Assay, Staining, Expressing, Marker, Immunofluorescence, Immunohistochemistry, Control

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: MIF-CD74 signaling coordinates epithelial-macrophage crosstalk in inflamed prostate tissue. (A) UMAP representation of single-cell RNA sequencing data from control (CTR) and EAP mouse prostates. (B) Sankey diagrams illustrating proportional changes in major and secondary cell populations, showing increased B cells, Th1(2) cells, and inflammatory macrophages (Inflam-Macro) in EAP. (C) CellChat analysis of human BPH single-cell transcriptomic data identifying monocytes/macrophages as the dominant signal-receiving population. (D) Within the MIF signaling pathway, epithelial cells act as the primary signal senders, whereas monocytes/macrophages represent the predominant recipients. (E) CellChat role analysis highlighting epithelial cells as major “Senders” and monocytes/macrophages as key “Receivers,” “Mediators,” and “Influencers” within the MIF network. (F) Heatmap summarizing the relative contribution of each cell type to the MIF signaling network based on role analysis. (G) Violin plots showing preferential expression of CD74 in monocytes/macrophages. (H) t-SNE feature plots of M1 module score, CD74, and CD68 expression, indicating enrichment of CD74 in M1-like macrophage subsets. (I–L) Spatial transcriptomic analysis using Cell2location revealing spatial co-localization of MIF-expressing epithelial cells (I–J) and CD74-expressing macrophages (K) within regions enriched for M1-like macrophages (L). Abbreviations: Abbreviations: CTR, control; EAP, experimental autoimmune prostatitis; UMAP, uniform manifold approximation and projection; Cell2location, spatial cell-type deconvolution tool; t-SNE, t-distributed stochastic neighbor embedding.

Article Snippet: Mouse primary prostate epithelial cells were obtained from Procell (Cat. No. CP-M064, Procell).

Techniques: RNA Sequencing, Control, Expressing

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: Inflammatory M1-like macrophages act as terminal effector cells driven by epithelial-derived MIF-CD74 signaling in chronic prostatitis. (A) UMAP plots of macrophage subtypes in CTR and EAP mouse prostates. (B) Frequencies of macrophage subpopulations, showing expansion of inflammatory macrophages (Inflam-Mac) in EAP. (C) AUCell-based scoring of M1-like polarization signatures across macrophage subsets. (D) AUCell-based scoring of M2-like polarization signatures across macrophage subsets. (E) KEGG pathway enrichment analysis of differentially expressed genes in macrophage subsets, indicating inflammatory and immunomodulatory programs. (F–G) Monocle pseudotime analysis revealing distinct trajectories, with C1qa expression declining and Nos2 / Il1b increasing during transition from proliferating to inflammatory macrophages. (H) Mif and Cd74 expression across CTR and EAP, showing upregulation in EAP. (I) Ridge plots of Cd74 expression across immune cell types, highlighting macrophage enrichment. (J) Multiplex immunofluorescence of mouse prostates showing co-localization of CD74 (green) and CD86 (red) in EAP versus CTR; nuclei stained with DAPI (blue). Scale bars, 50 μm. (K) Multiplex immunofluorescence of human BPH tissues showing increased CD74 − CD86 co-localization in regions with moderate/severe versus mild inflammation. Data are presented as the mean ± SD. ns, not significant; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. Abbreviations: CTR, control; EAP, experimental autoimmune prostatitis; UMAP, uniform manifold approximation and projection; AUCell, area under the curve for gene set activity; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Article Snippet: Mouse primary prostate epithelial cells were obtained from Procell (Cat. No. CP-M064, Procell).

Techniques: Derivative Assay, Expressing, Multiplex Assay, Immunofluorescence, Staining, Control, Activity Assay

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: Mouse primary prostate epithelial cells were obtained from Procell (Cat. No. CP-M064, Procell).

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

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: Schematic illustration of the epithelial ROS-ZNF24-MIF-CD74-PKM2-NF-κB signaling axis in chronic prostatitis. In response to epithelial injury or inflammatory stimuli, excessive ROS accumulate in prostatic epithelial cells, leading to the activation of the redox-responsive transcription factor ZNF24. Activated ZNF24 directly binds to the MIF promoter and drives MIF transcription, resulting in enhanced MIF production and release. Epithelial-derived MIF subsequently acts in a paracrine manner by binding to CD74 on macrophages. This interaction promotes the stabilization and nuclear translocation of PKM2, which acts as a co-activator to increase NF-κB signaling via p65 nuclear translocation. PKM2 activation also enhances glycolytic flux and contributes to mitochondrial dysfunction, further promoting M1 macrophage polarization and inflammatory cytokine production. Disruption of this axis via ISO-1, CD74 blockade, or PKM2 modulation (DASA-58) effectively mitigates chronic inflammation in the prostate.

Article Snippet: Mouse primary prostate epithelial cells were obtained from Procell (Cat. No. CP-M064, Procell).

Techniques: Activation Assay, Derivative Assay, Binding Assay, Translocation Assay, Disruption

Journal: Cancer Management and Research

Article Title: Normal Basal Epithelial Cells Stimulate the Migration and Invasion of Prostate Cancer Cell RM-1 by TGF-β1/STAT3 Axis in vitro

doi: 10.2147/CMAR.S303122

Figure Lengend Snippet: Identification of basal epithelial cells acquired from mouse prostate with anti-p63 in immunofluorescence assay (Objective 10X). Basal epithelial cells were specific to anti-p63 and more than 90%.

Article Snippet: Mouse prostate cancer cells RM-1 and mouse primary normal prostate basal epithelial cells were obtained from iCell Bioscience Inc. in Shanghai.

Techniques: Immunofluorescence

Journal: Cancer Management and Research

Article Title: Normal Basal Epithelial Cells Stimulate the Migration and Invasion of Prostate Cancer Cell RM-1 by TGF-β1/STAT3 Axis in vitro

doi: 10.2147/CMAR.S303122

Figure Lengend Snippet: Normal basal epithelial cells could stimulate the migration of RM-1 directly or indirectly. ( A ) Effect of basal epithelial cells on cells wound healing of RM-1 was determined after being mixed-cultured (1:1) for 0 h and 24 h. ( B ) Quantification of migrated width of wound healing assay. ( C ) Effect of basal epithelial cells on migration of RM-1 was determined after being co-cultured for 20 h. ( D ) Quantification of migrated cells in ( C ). ( E ) Effect of 10% conditional medium of basal epithelial cells on migration of RM-1 was estimated after being cultured for 20 h. ( F ) Quantification of migrated cells in (E). Values are represented by mean±SD from at least three independent experiments. “*” represents “p<0.05”, “***” represents “p<0.001” vs control group.

Article Snippet: Mouse prostate cancer cells RM-1 and mouse primary normal prostate basal epithelial cells were obtained from iCell Bioscience Inc. in Shanghai.

Techniques: Migration, Cell Culture, Wound Healing Assay, Control

Journal: Cancer Management and Research

Article Title: Normal Basal Epithelial Cells Stimulate the Migration and Invasion of Prostate Cancer Cell RM-1 by TGF-β1/STAT3 Axis in vitro

doi: 10.2147/CMAR.S303122

Figure Lengend Snippet: Normal basal epithelial cell promoted the invasion of RM-1 directly or indirectly. ( A ) Effect of basal epithelial cells on invasion of RM-1 was determined after being co-cultured for 20 h. ( B ) Quantification of invasive cells in ( A ). ( C ) Effect of 10% conditional medium of basal epithelial cells on invasion of RM-1 was estimated after being cultured for 20 h. ( D ) Quantification of invasive cells in ( C ). Values are represented by mean±SD from at least three independent experiments. “**” represents “p<0.01”, “***” represents “p<0.001” vs control group.

Article Snippet: Mouse prostate cancer cells RM-1 and mouse primary normal prostate basal epithelial cells were obtained from iCell Bioscience Inc. in Shanghai.

Techniques: Cell Culture, Control

Journal: Cancer Management and Research

Article Title: Normal Basal Epithelial Cells Stimulate the Migration and Invasion of Prostate Cancer Cell RM-1 by TGF-β1/STAT3 Axis in vitro

doi: 10.2147/CMAR.S303122

Figure Lengend Snippet: Basal epithelial cells secreted TGF-β1 and additional TGF-β1 stimulated the migration and invasion of RM-1. ( A ) Quantification of TGF-β1 levels measurement in ELISA assay. Effect of 5 ng/mL TGF-β1 on migration ( B ) and invasion ( C ) of RM-1 in transwell chamber with or without matrigel after being cultured for 20 h. ( D ) Quantification of invasive cells in ( C ). Values are represented by mean±SD from at least three independent experiments. “***” represents “p<0.001” vs control group.

Article Snippet: Mouse prostate cancer cells RM-1 and mouse primary normal prostate basal epithelial cells were obtained from iCell Bioscience Inc. in Shanghai.

Techniques: Migration, Enzyme-linked Immunosorbent Assay, Cell Culture, Control

Journal: Cancer Management and Research

Article Title: Normal Basal Epithelial Cells Stimulate the Migration and Invasion of Prostate Cancer Cell RM-1 by TGF-β1/STAT3 Axis in vitro

doi: 10.2147/CMAR.S303122

Figure Lengend Snippet: Basal epithelial cells secreted TGF-β1 to promote the migration and invasion of RM-1. ( A ) Basal epithelial cells and TGF-β1 (5 ng/mL) changed the morphology of RM-1 from cuboid to spindle-shape. ( B ) HE staining and immunocytochemistry with anti-63 in mixed-cultured group. Arrowhead indicates basal epithelial cell of which p63 is positive. ( C ) Basal epithelial cells and TGF-β1 regulated the proteins associated with EMT. ( D ) Quantification of relative proteins expression in ( C ). ( E ) Effect of different concentration of SB431542 on RM-1 death was measured by CCK8 assay. SB431542 (2000 nM, 4000 nM) inhibited the migration ( F and G ) and invasion ( H and I ) of co-cultured RM-1. ( J ) Expressions of EMT relevant proteins were down-regulated when SB431542 (2000 nM or 4000 nM) was added into medium of co-cultured cells. ( K ) Quantification of relative expressions of proteins in ( J ). Values are represented by mean±SD from at least three independent experiments. “*” represents “p<0.05”. “**” represents “p<0.01”. “***” represents “p<0.001”.

Article Snippet: Mouse prostate cancer cells RM-1 and mouse primary normal prostate basal epithelial cells were obtained from iCell Bioscience Inc. in Shanghai.

Techniques: Migration, Staining, Immunocytochemistry, Cell Culture, Expressing, Concentration Assay, CCK-8 Assay

Journal: Cancer Management and Research

Article Title: Normal Basal Epithelial Cells Stimulate the Migration and Invasion of Prostate Cancer Cell RM-1 by TGF-β1/STAT3 Axis in vitro

doi: 10.2147/CMAR.S303122

Figure Lengend Snippet: TGF-β1 which was secreted by basal epithelial cells could promote migration and invasion of RM-1 depending on the phosphorylation of STAT3. ( A and B ) p-STAT3 was up-regulated by basal epithelial cells, conditional medium of basal epithelial cells and TGF-β1. ( C and D ) Expression of p-STAT3 in co-cultured group was down-regulated when SB431542 was added. ( E ) Stattic down-regulated the expression of p-STAT3 and STAT3. Stattic inhibited the proliferation ( F ), migration ( G ) and invasion ( H ) of untreated RM-1 and co-cultured RM-1. Values are represented by mean±SD from at least three independent experiments. “**” represents “p<0.01”. “***” represents “p<0.001”.

Article Snippet: Mouse prostate cancer cells RM-1 and mouse primary normal prostate basal epithelial cells were obtained from iCell Bioscience Inc. in Shanghai.

Techniques: Migration, Phospho-proteomics, Expressing, Cell Culture