Journal: Frontiers in Immunology

Article Title: Identify GDPD3 as a key regulator of epithelial–mesenchymal transition and prostate adenocarcinoma progression via the LPA/LPAR1/AKT axis: transcriptomic and experimental study

doi: 10.3389/fimmu.2025.1637325

Figure Lengend Snippet: GDPD3 promotes EMT through LPA–LPAR1 signaling. (A) Analysis of the correlation between GDPD3 and LPA. (B) LPA levels were measured by ELISA following GDPD3 overexpression in DU145 cells. (C) DU145 cells were treated with either vehicle control or 20 μM LPA for 24, 48, and 72 hours. Protein levels of N-cadherin, E-cadherin and Vimentin were assessed by Western blot and quantified. (D, E) DU145 cells were treated with LPA alone, or with LPA in combination with KI6425 or AM095 (both LPAR1 inhibitors). EMT markers N-cadherin, Vimentin and α-SMA were evaluated by Western blot, and band intensities were quantified using ImageJ software, normalized to actin. (n = 3–4 independent experiments). Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The DU145 human prostate cancer cell line was obtained from the American Type Culture Collection (ATCC) in February 2025.

Techniques: Enzyme-linked Immunosorbent Assay, Over Expression, Control, Western Blot, Software

Journal: Frontiers in Immunology

Article Title: Identify GDPD3 as a key regulator of epithelial–mesenchymal transition and prostate adenocarcinoma progression via the LPA/LPAR1/AKT axis: transcriptomic and experimental study

doi: 10.3389/fimmu.2025.1637325

Figure Lengend Snippet: LPAR1 knockdown and AKT inhibition attenuate LPA-induced EMT. (A, B) LPAR1 was knocked down, and knockdown efficiency was subsequently validated. (C, D) DU145 cells were transfected with siRNA targeting LPAR1, and the expression levels of N-cadherin, E-cadherin and Vimentin were assessed by Western blot. (E, F) Western blot analysis was performed to assess the expression levels of AKT and phosphorylated AKT (p-AKT). (G, H) DU145 cells were treated with 2 μM LY294002 (AKT pathway inhibitor), and the protein levels of AKT, phosphorylated AKT (p-AKT), N-cadherin, E-cadherin and Vimentin were assessed by Western blot and quantified. Band intensities were quantified using ImageJ software and normalized to actin. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The DU145 human prostate cancer cell line was obtained from the American Type Culture Collection (ATCC) in February 2025.

Techniques: Knockdown, Inhibition, Transfection, Expressing, Western Blot, Software

Journal: Biochemistry and Biophysics Reports

Article Title: Investigating the ABTS-based antioxidant potential and antiproliferative activity of pulp, skin and seeds extracts from Nephelium lappaceum , Manilkara zapota , and Meliccocus oliviformis on human prostate and colon cancer cells

doi: 10.1016/j.bbrep.2025.102257

Figure Lengend Snippet: Effects of the different fruit extracts on the proliferation of the colon cancer cell line HCT116 and the prostate cancer cell line DU145 . DU145 (A) or HCT116 (B) were seeded in a 12-well plate and allowed to grow for 12 h. Then, 500 μg/mLof PBS-resuspended extract derived from the pulp, seeds, and skin of Meliccocus oliviformis , Nephelium lappaceum , and Manilkara zapota were added and cell proliferation was monitored by analyzing the surface area occupied by the cells in each well (% confluence) from photos taken at regular intervals (every 3 h) by the Incucyte live cell imaging system over a period of three days. Data shown are the average ± SD of three independent experiments.

Article Snippet: The HCT116 (colic), and DU145 (prostatic) human cancer cell lines were purchased from ATCC (American Type Culture Collection).

Techniques: Derivative Assay, Live Cell Imaging

Journal: Biochemistry and Biophysics Reports

Article Title: Investigating the ABTS-based antioxidant potential and antiproliferative activity of pulp, skin and seeds extracts from Nephelium lappaceum , Manilkara zapota , and Meliccocus oliviformis on human prostate and colon cancer cells

doi: 10.1016/j.bbrep.2025.102257

Figure Lengend Snippet: Effects of the different fruit extracts on the viability of the colon cancer cell line HCT116 and the prostate cancer cell line DU145 . DU145 (A) or HCT116 (B) were seeded in a 12-well plate and allowed to grow for 12 h. Then, 500 μg/mL of PBS-resuspended extract derived from the pulp, seeds, and skin of Meliccocus oliviformis , Nephelium lappaceum , and Manilkara zapota were added. 24h later, cell viability was assessed using the MTT assay. Data shown are the average ± SD of three independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, one-way ANOVA followed by Tukey's multiple comparison test.

Article Snippet: The HCT116 (colic), and DU145 (prostatic) human cancer cell lines were purchased from ATCC (American Type Culture Collection).

Techniques: Derivative Assay, MTT Assay, Comparison

Journal: Antioxidants

Article Title: PTEN/PKM2/ERα-Driven Glyoxalase 1 Overexpression Sustains PC3 Prostate Cancer Cell Growth Through MG-H1/RAGE Pathway Desensitization Leading to H 2 O 2 -Dependent KRIT1 Downregulation

doi: 10.3390/antiox14091120

Figure Lengend Snippet: Glyoxalase 1 (Glo1) is overexpressed in aggressive prostate cancer (PCa) tissues and cell lines. Representative images of Glo1 protein expression, evaluated by immunohistochemistry, in PCa tissues from ( a ) low-grade (LG) and ( b ) high-grade (HG) patients with PCa. Magnification 200×. ( c ) Glo1 protein expression, measured by ELISA; ( d ) Glo1 specific enzyme activity, evaluated by a specific spectrophotometric method; ( e ) cell proliferation, measured by the CCK-8 assay and the clonogenic assay; ( f ) 20 µM etoposide and 1 µM staurosporine-driven apoptosis, evaluated by the increased expression of active caspase-3, using ELISA; and ( g ) MG-H1 intracellular amounts, measured by a specific ELISA kit, in moderately aggressive DU145 and highly aggressive PC3 PCa cells. The histograms indicate the mean ± SD of three different cultures. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human prostate cancer cell lines DU145 (derived from brain metastasis) and PC3 (derived from bone metastasis) were acquired from the American Type Culture Collection (ATCC) and maintained, following the supplier’s instructions, at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Expressing, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Activity Assay, CCK-8 Assay, Clonogenic Assay

Journal: Antioxidants

Article Title: PTEN/PKM2/ERα-Driven Glyoxalase 1 Overexpression Sustains PC3 Prostate Cancer Cell Growth Through MG-H1/RAGE Pathway Desensitization Leading to H 2 O 2 -Dependent KRIT1 Downregulation

doi: 10.3390/antiox14091120

Figure Lengend Snippet: Glyoxalase 1 (Glo1) sustains prostate cancer (PCa) aggressiveness to control methylglyoxal (MG)-derived hydroimidazolone 1 (MG-H1). ( a ) MG-H1 intracellular amounts, measured by a specific ELISA kit, and ( b ) cell proliferation, measured by the CCK-8 assay, of PC3 cells upon Glo1 transient silencing (siGlo1) ( a , b ) and of DU145 cells upon Glo1 ectopic expression (pCMV-Glo1) ( a , b ). The histograms indicate the mean ± SD of three different cultures. siCtr: control (non-specific siRNA); pCMV-Ctr: control containing plasmid DNA. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human prostate cancer cell lines DU145 (derived from brain metastasis) and PC3 (derived from bone metastasis) were acquired from the American Type Culture Collection (ATCC) and maintained, following the supplier’s instructions, at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Control, Derivative Assay, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Expressing, Plasmid Preparation

Journal: Antioxidants

Article Title: PTEN/PKM2/ERα-Driven Glyoxalase 1 Overexpression Sustains PC3 Prostate Cancer Cell Growth Through MG-H1/RAGE Pathway Desensitization Leading to H 2 O 2 -Dependent KRIT1 Downregulation

doi: 10.3390/antiox14091120

Figure Lengend Snippet: Glyoxalase 1 (Glo1) upregulation is driven by the PTEN-dependent pathway. ( a ) PTEN expression, measured by a specific ELISA kit, in PC3 cells upon PTEN ectopic expression (pCMV-PTEN) decreased ( b ) Glo1 mRNA expression and specific enzyme activity, evaluated by qRT-PCR and a specific spectrophotometric method, respectively; ( c ) PTEN expression, measured by a specific ELISA kit, in transiently PTEN-knocked-down (siPTEN) DU145 cells increased ( d ) Glo1 transcript levels and specific enzyme activity. The histograms indicate the mean ± SD of three different cultures. pCMV-Ctr: control containing plasmid DNA; siCtr: control (non-specific siRNA). *** p < 0.001, **** p < 0.0001.

Article Snippet: Human prostate cancer cell lines DU145 (derived from brain metastasis) and PC3 (derived from bone metastasis) were acquired from the American Type Culture Collection (ATCC) and maintained, following the supplier’s instructions, at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Activity Assay, Quantitative RT-PCR, Control, Plasmid Preparation

Journal: Antioxidants

Article Title: PTEN/PKM2/ERα-Driven Glyoxalase 1 Overexpression Sustains PC3 Prostate Cancer Cell Growth Through MG-H1/RAGE Pathway Desensitization Leading to H 2 O 2 -Dependent KRIT1 Downregulation

doi: 10.3390/antiox14091120

Figure Lengend Snippet: Glyoxalase 1 (Glo1) upregulation is driven by the PI3/AKT/mTOR pathway. ( a , c ) Ectopic expression of PTEN (pCMV-PTEN) in PC3 cells or ( b , d ) PTEN knockdown (siPTEN) in DU145 cells led to desensitization or activation of the PI3K/AKT/mTOR pathway, respectively. Treatment of PC3 cells with the PI3K inhibitor LY294002 (LY), the AKT inhibitor MK2206 (MK), and the mTOR inhibitor rapamycin (Rapam) further confirmed the involvement of the PI3K/AKT/mTOR signaling cascade in regulating ( e ) Glo1 at both the transcript and the functional level. The PI3/AKT/mTOR pathway was evaluated by p-AKT and p-mTOR expression measured by both specific ELISA kits and Western blotting. mRNA expression was evaluated by qRT-PCR, while specific enzyme activity was measured by a specific spectrophotometric method. β-actin served as the internal control. The histograms indicate the mean ± SD of three different cultures. pCMV-Ctr: control containing plasmid DNA; siCtr: control (non-specific siRNA). ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Human prostate cancer cell lines DU145 (derived from brain metastasis) and PC3 (derived from bone metastasis) were acquired from the American Type Culture Collection (ATCC) and maintained, following the supplier’s instructions, at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Expressing, Knockdown, Activation Assay, Functional Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Quantitative RT-PCR, Activity Assay, Control, Plasmid Preparation

Journal: Antioxidants

Article Title: PTEN/PKM2/ERα-Driven Glyoxalase 1 Overexpression Sustains PC3 Prostate Cancer Cell Growth Through MG-H1/RAGE Pathway Desensitization Leading to H 2 O 2 -Dependent KRIT1 Downregulation

doi: 10.3390/antiox14091120

Figure Lengend Snippet: Expression of the receptor for AGEs (RAGE), levels of hydrogen peroxide (H 2 O 2 ), and KRIT1 expression in DU145 and PC3 cells. ( a ) RAGE transcript levels, measured by qRT-PCR, and RAGE protein expression, evaluated by Western blotting. ( b ) H 2 O 2 levels were measured by a specific kit, and ( c ) KRIT1 expression was studied by using a specific ELISA kit. β-actin served as the internal control. The histograms represent the mean ± SD from three independent experiments. ** p < 0.01, *** p < 0.001.

Article Snippet: Human prostate cancer cell lines DU145 (derived from brain metastasis) and PC3 (derived from bone metastasis) were acquired from the American Type Culture Collection (ATCC) and maintained, following the supplier’s instructions, at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Control

Journal: Cell Communication and Signaling : CCS

Article Title: ACACA depletion activates the cPLA2–arachidonic acid–NF-κB axis to drive inflammatory reprogramming in androgen receptor-independent prostate cancer

doi: 10.1186/s12964-025-02363-0

Figure Lengend Snippet: Depletion of ACACA promotes inflammation in ARIPC cells. ( A-B ) Inflammatory pathways significantly enriched in ACACA-depleted ARIPC cells. ( C-D ) GSEA showed that the ‘Regulation of inflammatory response’ was activated in ACACA-depleted ARIPC cells on the basis of the transcriptome data. ( E-F ) Heatmap of inflammatory biomarker-related gene expression in ACACA-depleted ARIPC cells on the basis of transcriptome data. ( G ) Quantification of the qPCR analysis results of inflammation biomarker-related gene expression in ACACA depleted DU145 cells. ( H ) Quantification of qPCR analysis results of inflammation biomarker-related gene expression in DU145 cells after TOFA treatment (10 µg/mL, 72 h). Data are represented as mean ± SD in ( G and H ). * P < 0.05

Article Snippet: The human androgen receptor-independent prostate cancer (ARIPC) cell lines DU145 and PC3, the androgen receptor positive prostate cancer cell line LNCaP, the benign prostatic hyperplasia cell line BPH1, and HEK293T cells were obtained from the American Type Culture Collection (ATCC, Manassas, USA).

Techniques: Biomarker Discovery, Gene Expression

Journal: Cell Communication and Signaling : CCS

Article Title: ACACA depletion activates the cPLA2–arachidonic acid–NF-κB axis to drive inflammatory reprogramming in androgen receptor-independent prostate cancer

doi: 10.1186/s12964-025-02363-0

Figure Lengend Snippet: ACACA depletion activates NF-κB signaling in ARIPC cells. ( A ) Western blot analysis of key proteins of the NF-κB signaling pathway and inflammatory signaling in ACACA-depleted DU145 cells. ( B-C ) Western blot analysis of P65 and pP65 after treatment with TOFA for 72 h at the indicated concentrations (µg/mL) in the indicated ARIPC cells. ( D ) and ( E ) Western blot analysis of P65 and pP65 expression following ACACA silencing in the indicated ARIPC cells. ( F-I ) Immunofluorescence analysis of P65 in ACACA-depleted DU145 cells generated with different shRNA sequences (shACACA#1 and shACACA#2), with the quantification presented in ( G ) and ( I ), respectively. ( J-K ) Immunofluorescence analysis of P65 in DU145 cells after TOFA treatment (72 h), with quantification presented in ( K ). ( L ) Western blot analysis of P65 and pP65 and ( M ) qPCR analysis of P65 after arachidonic acid (AA) treatment (50 µM, 72 h). ( N ) Western blot analysis of P65 and pP65 and ( M ) qPCR analysis of P65 after cPLA2 silencing in ACACA-depleted DU145 cells. ( O ) qPCR analysis of P65. Data are presented as the means ± SD in ( G, I, K, M, O ). * P < 0.05

Article Snippet: The human androgen receptor-independent prostate cancer (ARIPC) cell lines DU145 and PC3, the androgen receptor positive prostate cancer cell line LNCaP, the benign prostatic hyperplasia cell line BPH1, and HEK293T cells were obtained from the American Type Culture Collection (ATCC, Manassas, USA).

Techniques: Western Blot, Expressing, Immunofluorescence, Generated, shRNA

Journal: Cell Communication and Signaling : CCS

Article Title: ACACA depletion activates the cPLA2–arachidonic acid–NF-κB axis to drive inflammatory reprogramming in androgen receptor-independent prostate cancer

doi: 10.1186/s12964-025-02363-0

Figure Lengend Snippet: Depletion of ACACA causes increased arachidonic acid activity in prostate cancer cells. ( A ) Levels of two metabolites associated with arachidonic acid from metabolomic analysis data after ACACA knockdown in DU145 cells. ( B-M ) Heatmap and individual panels of arachidonic acid and its metabolite levels in ACACA-depleted PC3 cells. ( N-O ) Heatmaps showing the transcriptional expression levels of arachidonic acid pathway metabolic enzymes in the indicated ARIPC cells. ( P-Q ) Indicated pathways significantly enriched in ACACA-depleted DU145 cells. Data are represented as mean ± SD in ( A , C-M ). * P < 0.05

Article Snippet: The human androgen receptor-independent prostate cancer (ARIPC) cell lines DU145 and PC3, the androgen receptor positive prostate cancer cell line LNCaP, the benign prostatic hyperplasia cell line BPH1, and HEK293T cells were obtained from the American Type Culture Collection (ATCC, Manassas, USA).

Techniques: Activity Assay, Knockdown, Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: ACACA depletion activates the cPLA2–arachidonic acid–NF-κB axis to drive inflammatory reprogramming in androgen receptor-independent prostate cancer

doi: 10.1186/s12964-025-02363-0

Figure Lengend Snippet: cPLA2-driven arachidonic acid (AA) promotes inflammation and migration in ACACA-depleted ARIPC cells. ( A ) qPCR analysis of inflammation biomarker-related gene expression in ACACA-depleted DU145 cells after 72 h of treatment with 50 µM AA. ( B-E ) Evaluation of the migration potential of ACACA-depleted DU145 cells following AA treatment (72 h) at the indicated concentrations, via wound healing ( B ) and Transwell assays ( D ), with the quantification presented in ( C ) and ( E ), respectively. ( F ) Transcriptome data and ( G ) Western blot analysis of cPLA2 expression in ACACA-depleted DU145 cells. ( H ) Correlation analysis between cPLA2 and ACACA expression levels in the SU2C PolyA cohort. ( I ) Violin plots illustrating cPLA2 expression levels in mCRPC samples divided into high and low ACACA expression groups in the SU2C PolyA cohort. ( J ) qPCR analysis of cPLA2 expression after 72 h of treatment with silencing RNA in ACACA-depleted DU145 cells. ( K ) AA concentration measured by ELISA following cPLA2 silencing for 72 h in ACACA-depleted DU145 cells. ( L ) qPCR analysis of inflammation biomarker-related gene expression after cPLA2 silencing in ACACA-depleted DU145 cells. ( M-P ) Evaluation of the migration potential of ACACA-depleted ARIPC cells after cPLA2 silencing via wound healing ( M ) and Transwell ( O ) assays, with the quantification presented in ( N ) and ( P ), respectively. The data are presented as the means ± SDs in A, C, E, F, I, J-L, N , and P. * P < 0.05

Article Snippet: The human androgen receptor-independent prostate cancer (ARIPC) cell lines DU145 and PC3, the androgen receptor positive prostate cancer cell line LNCaP, the benign prostatic hyperplasia cell line BPH1, and HEK293T cells were obtained from the American Type Culture Collection (ATCC, Manassas, USA).

Techniques: Migration, Biomarker Discovery, Gene Expression, Western Blot, Expressing, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Cell Communication and Signaling : CCS

Article Title: ACACA depletion activates the cPLA2–arachidonic acid–NF-κB axis to drive inflammatory reprogramming in androgen receptor-independent prostate cancer

doi: 10.1186/s12964-025-02363-0

Figure Lengend Snippet: The NF-κB signaling pathway was enriched in ACACA-depleted ARIPC cells. ( A-B ) GSEA-derived significant pathways in KEGG signaling transduction identified from transcriptome data of ACACA-depleted ARIPC cells. ( C ) Venn diagram of KEGG signaling transduction pathways in ACACA-depleted DU145 and PC3 cells. The intersection indicates the number of shared pathways. ( D-E ) Heatmap of the expression of key genes associated with the intersecting pathways in ( C ). ( F ) Venn diagram illustrating the overlapping genes between the DU145 ( D ) and PC3 ( E ) cell lines. ( G-H ) NF-κB-related pathways significantly enriched in ACACA-depleted ARIPC cells. ( I ) UMAP plots of single-cell RNA sequencing (scRNA-seq) data from 6 metastatic prostate cancer patients after ARSI treatment ( GSE137829 ), with luminal epithelial cells highlighted by a black circle for subsequent analysis. ( J ) UMAP plots of luminal epithelial cells in ( I ) clustered into AR-positive and AR-negative subpopulations on the basis of the GSVA score. ( K ) UMAP plots of AR-negative cells in ( J ), which were further clustered into ACACA-high ( n = 91) and ACACA-low ( n = 285) subpopulations on the basis of mRNA expression. ( L ) GO enrichment analysis of DEGs between the ACACA-high and ACACA-low subpopulations in ( K ) revealed significant enrichment of the NF-κB and TNF signaling pathways

Article Snippet: The human androgen receptor-independent prostate cancer (ARIPC) cell lines DU145 and PC3, the androgen receptor positive prostate cancer cell line LNCaP, the benign prostatic hyperplasia cell line BPH1, and HEK293T cells were obtained from the American Type Culture Collection (ATCC, Manassas, USA).

Techniques: Derivative Assay, Transduction, Expressing, RNA Sequencing, Protein-Protein interactions

Journal: Cell Communication and Signaling : CCS

Article Title: ACACA depletion activates the cPLA2–arachidonic acid–NF-κB axis to drive inflammatory reprogramming in androgen receptor-independent prostate cancer

doi: 10.1186/s12964-025-02363-0

Figure Lengend Snippet: Inhibition of NF-κB suppresses inflammation and migration in ACACA-depleted ARIPC cells. ( A ) Western blot analysis of key proteins in the NF-κB signaling pathway and inflammatory signaling pathway after treatment with QNZ (10 nM, 72 h) in ACACA-depleted DU145 cells. ( B ) qPCR analysis of inflammation-related genes after treatment with QNZ (10 nM, 72 h) in ACACA-depleted DU145 cells. ( C-F ) Evaluation of the migration potential of ACACA-depleted ARIPC cells following treatment with QNZ (10 nM, 72 h) via wound healing ( C ) and Transwell ( E ) assays, with the quantification presented in ( D ) and ( F ), respectively. ( G ) qPCR analysis of Slug and Sanil in ACACA-depleted DU145 cells. ( H ) qPCR analysis of Slug and Sanil in the ACACA-depleted DU145 cells after treatment with QNZ (10 nM, 72 h). The data are presented as the means ± SD in ( B, D, F, G, H ). * P < 0.05

Article Snippet: The human androgen receptor-independent prostate cancer (ARIPC) cell lines DU145 and PC3, the androgen receptor positive prostate cancer cell line LNCaP, the benign prostatic hyperplasia cell line BPH1, and HEK293T cells were obtained from the American Type Culture Collection (ATCC, Manassas, USA).

Techniques: Inhibition, Migration, Western Blot