Journal: Oncotarget

Article Title: The metastasis suppressor CD82/KAI1 inhibits fibronectin adhesion-induced epithelial-to-mesenchymal transition in prostate cancer cells by repressing the associated integrin signaling

doi: 10.18632/oncotarget.13767

Figure Lengend Snippet: A. Human prostate epithelial (PZ-HPV-7) and cancer (DU145, LNCaP, and PC3) cell lines and stably CD82-transfected DU145 and antisense CD82 fragment-transfected LNCaP cell clones were assessed for CD82 expression levels through immunoblotting analysis. B. Stably CD82-transfected DU145 and antisense CD82 fragment-transfected LNCaP cell clones grown on fibronectin (FN) were viewed under a phase-contrast microscope. Scale bar, 10 μm. C. The cells were seeded onto plates precoated with poly-L(+)-lysine (p-Lys) or FN and cultured for the indicated time periods. Expression of E-cadherin and mesenchymal marker proteins was examined through immunoblotting analysis using antibodies specific to each protein.

Article Snippet: PZ-HPV-7 (ATCC), a human prostate epithelial cell line transformed by human papillomavirus-18, was cultured in K-SFM medium (Invitrogen, Carlsbad, CA) supplemented with 0.05 mg/ml bovine pituitary extract and 5 ng/ml EGF.

Techniques: Stable Transfection, Transfection, Clone Assay, Expressing, Western Blot, Microscopy, Cell Culture, Marker

Journal: Oncotarget

Article Title: The metastasis suppressor CD82/KAI1 inhibits fibronectin adhesion-induced epithelial-to-mesenchymal transition in prostate cancer cells by repressing the associated integrin signaling

doi: 10.18632/oncotarget.13767

Figure Lengend Snippet: A. PZ-HPV-7 prostate epithelial cells were lysed with Brij 97 detergent, and immunoprecipitation (IP) was performed with normal mouse IgG or anti-CD82 antibody. The immunoprecipitates were analyzed by immnublotting using anti-integrin β 1 , α 3 , α 5 , or α 6 antibody. B. CD82 mutant cDNA, which encodes CD82 with a large extracellular loop (LEL) substituted with that of TM4SF2 as illustrated, was generated by PCR and subcloned into the pAdEasy-1 adenoviral vector to produce recombinant adenovirus. C. CD82-deficient PC3 prostate cancer cells grown on fibronectin (FN) were infected with adenovirus containing a wild-type (wt) or mutant (mt) CD82 expression construct, and Brij 97 detergent lysates were subjected to immunoprecipitation with an anti-β 1 integrin antibody followed by immunoblotting analysis using antibodies that recognize the C-terminus or LEL of CD82 and the LEL of TM4SF2. D. PC3 cells grown on poly-L(+)-lysine (p-Lys) or FN were infected with adenovirus containing a wt- or mt-CD82 expression construct and then assessed for the protein levels of E-cadherin and Snail. E. PC3 cells grown on FN were infected with wt-CD82 construct-containing adenovirus either alone or together with mt-CD82 construct-containing adenovirus and examined for E-cadherin and Snail expression. Numbers in parentheses represent the MOI values of adenovirus.

Article Snippet: PZ-HPV-7 (ATCC), a human prostate epithelial cell line transformed by human papillomavirus-18, was cultured in K-SFM medium (Invitrogen, Carlsbad, CA) supplemented with 0.05 mg/ml bovine pituitary extract and 5 ng/ml EGF.

Techniques: Immunoprecipitation, Mutagenesis, Generated, Plasmid Preparation, Recombinant, Infection, Expressing, Construct, Western Blot

Journal: BMC Cancer

Article Title: The combination of poly(ADP-ribose) polymerase inhibitor and statin inhibits the proliferation of human castration-resistant and taxane-resistant prostate cancer cells in vitro and in vivo

doi: 10.1186/s12885-025-13895-6

Figure Lengend Snippet: Effect of simvastatin on DNA repair gene expression in prostate cancer and prostate stromal cells (PrSC). PC-3, 22Rv1, LNCaP-LA, LNCaP, and PrSC cells were incubated with the medium containing 10% FBS for 24 h, and the medium was switched to the indicated concentration of simvastatin in the medium containing 10% FBS. After 48 h, the total RNA ( A , C ) and total protein ( B , D ) were collected. ( A ) Comparison of mRNA expression levels of each gene in each cell without simvastatin. mRNA expression of BRCA1, BRCA2, RAD51, FANCD2, FANCG, FANCA, BARD1, RFC3, RFC4, and RFC5 was evaluated by performing real-time polymerase chain reaction tests, and the relative quantitative volume (RQV) was calculated by comparing the expression of β-actin. Values are expressed as mean ± standard deviation (SD) ( n = 3). * P < 0.05 vs. the other cells. ( B ) Comparison of protein levels of BRCA1 and RAD51 in each cell without simvastatin. The protein expression of BRCA1 and RAD51 was evaluated by western blotting. A representative experiment is shown, which was repeated three times with similar results. ( C ) Comparison of mRNA expression levels of each gene in each cell after treatment of simvastatin. mRNA expression of BRCA1, BRCA2, RAD51, FANCD2, FANCG, FANCA, BARD1, RFC3, RFC4, and RFC5 was evaluated by performing real-time polymerase chain reaction tests, and the relative quantitative volume (RQV) was calculated by comparing the expression of β-actin. Values are expressed as mean ± standard deviation (SD) ( n = 3). * P < 0.05 vs. 0 µM of each cell. ( D ) Comparison of BRCA1 protein levels in each cell after treatment of simvastatin. The protein expression of BRCA1 was evaluated by western blotting. A representative experiment is shown, which was repeated three times with similar results

Article Snippet: Human prostate cancer cell lines PC-3, LNCaP, and 22Rv1 were purchased from DS Pharma Biomedical (Osaka, Japan) and cultured in RPMI 1640 (Sigma-Aldrich, St. Louis, MO) supplemented with 10% fetal bovine serum (FBS) (Moregate, Bulimba, Australia).

Techniques: Gene Expression, Incubation, Concentration Assay, Comparison, Expressing, Real-time Polymerase Chain Reaction, Standard Deviation, Western Blot

Journal: BMC Cancer

Article Title: The combination of poly(ADP-ribose) polymerase inhibitor and statin inhibits the proliferation of human castration-resistant and taxane-resistant prostate cancer cells in vitro and in vivo

doi: 10.1186/s12885-025-13895-6

Figure Lengend Snippet: Effect of combination therapy with simvastatin and olaparib on androgen-independent prostate cancer cells. The cells were then incubated in a medium. After 48 h, the cells were cultured in a medium with or without simvastatin (PC-3, 2 µM; other cells, 5 µM) and olaparib (10 µM). After 72 h for PC-3, LNCaP, and PrSC and 120 h for LNCaP-LA and 22Rv1 cells, the number of viable cells was evaluated using MTS assay ( A ) and cell counts ( B ). Values are expressed as mean ± standard deviation (SD) (A; n = 5, B; n = 4). * p < 0.05. Sim; simvastatin, Ola; olaparib

Article Snippet: Human prostate cancer cell lines PC-3, LNCaP, and 22Rv1 were purchased from DS Pharma Biomedical (Osaka, Japan) and cultured in RPMI 1640 (Sigma-Aldrich, St. Louis, MO) supplemented with 10% fetal bovine serum (FBS) (Moregate, Bulimba, Australia).

Techniques: Incubation, Cell Culture, MTS Assay, Standard Deviation

Journal: BMC Cancer

Article Title: The combination of poly(ADP-ribose) polymerase inhibitor and statin inhibits the proliferation of human castration-resistant and taxane-resistant prostate cancer cells in vitro and in vivo

doi: 10.1186/s12885-025-13895-6

Figure Lengend Snippet: Effect of combination therapy with simvastatin and olaparib on tumor growth and BRCA1 expression in vivo in a 22Rv1 xenograft model. Graphic view of mean tumor volumes in 22Rv1 xenografts. Mice were injected with 22Rv1 cells (3 × 10 6 per site) and subsequently orally treated with simvastatin (100 mg/kg once daily) with or without olaparib (50 mg/kg once daily). Line graphs, mean ( n = 5); bars, standard deviation (SD); * p < 0.05 vs. the other group. BRCA1 mRNA expression in xenograft tissue was analyzed via quantitative real-time PCR (B). Values are expressed as mean ± SD ( n = 5). Sim; simvastatin, Ola; olaparib

Article Snippet: Human prostate cancer cell lines PC-3, LNCaP, and 22Rv1 were purchased from DS Pharma Biomedical (Osaka, Japan) and cultured in RPMI 1640 (Sigma-Aldrich, St. Louis, MO) supplemented with 10% fetal bovine serum (FBS) (Moregate, Bulimba, Australia).

Techniques: Expressing, In Vivo, Injection, Standard Deviation, Real-time Polymerase Chain Reaction

Journal: BMC Cancer

Article Title: The combination of poly(ADP-ribose) polymerase inhibitor and statin inhibits the proliferation of human castration-resistant and taxane-resistant prostate cancer cells in vitro and in vivo

doi: 10.1186/s12885-025-13895-6

Figure Lengend Snippet: Effect of combination therapy with simvastatin and olaparib on taxane-resistant prostate cancer cells. A and B . Comparison of DNA repair gene expression in 22Rv1 and 22Rv1-CR cells. A . mRNA expression of BRCA1, BRCA2, RAD51, BARD1, and FANCA was evaluated by RT-PCR, and the relative quantitative volume was calculated by comparing the expression of β-actin. Values are expressed as means ± standard deviations (SD) ( n = 3). * P < 0.05 vs. 22Rv1 cells. B . Protein expression of BRCA1, RAD51, BARD1, and FANCA was evaluated using western blotting. A representative experiment is shown, which was repeated three times with similar results. C and D . Cells were incubated in medium containing various concentrations of simvastatin. After 48 h, the total RNA ( C ) and total protein ( D ) were collected. A . mRNA expression of BRCA1, BRCA2, RAD51, BARD1, and FANCA was evaluated by RT-PCR, and the relative quantitative volume was calculated by comparing the expression of β-actin. Values are expressed as mean ± standard deviation (SD) ( n = 3). * P < 0.05 vs. 0 µM of each gene. D . Protein expression of BRCA1 and FANCA was evaluated by western blotting. A representative experiment is shown, which was repeated three times with similar results. E and F. Cells were incubated in the medium. After 48 h, the cells were cultured in a medium containing olaparib (10 µM) with or without simvastatin (5 µM). After 96 h, the number of viable cells was evaluated using the MTS assay ( E ) and cell counts ( F ). * P < 0.05. Sim; simvastatin, Ola; olaparib

Article Snippet: Human prostate cancer cell lines PC-3, LNCaP, and 22Rv1 were purchased from DS Pharma Biomedical (Osaka, Japan) and cultured in RPMI 1640 (Sigma-Aldrich, St. Louis, MO) supplemented with 10% fetal bovine serum (FBS) (Moregate, Bulimba, Australia).

Techniques: Comparison, Gene Expression, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Incubation, Standard Deviation, Cell Culture, MTS Assay

Journal: BMC Cancer

Article Title: The combination of poly(ADP-ribose) polymerase inhibitor and statin inhibits the proliferation of human castration-resistant and taxane-resistant prostate cancer cells in vitro and in vivo

doi: 10.1186/s12885-025-13895-6

Figure Lengend Snippet: Effect of combination therapy with simvastatin and olaparib on γΗ2ΑΧ expression in androgen-independent or taxane-resistant prostate cancer cells. Cells were incubated in a medium. After 48 h, cells were cultured in a medium with or without simvastatin (PC-3; 2 µM, other cells; 5 µM) and olaparib (10 µM). Proteins were collected after 48 h for PC-3 and LNCaP-LA cells and 96 h for 22Rv1 and 22Rv1-CR cells. Protein expression of γΗ2ΑΧ was evaluated using western blotting. A representative experiment is shown, which was repeated three times with similar results

Article Snippet: Human prostate cancer cell lines PC-3, LNCaP, and 22Rv1 were purchased from DS Pharma Biomedical (Osaka, Japan) and cultured in RPMI 1640 (Sigma-Aldrich, St. Louis, MO) supplemented with 10% fetal bovine serum (FBS) (Moregate, Bulimba, Australia).

Techniques: Expressing, Incubation, Cell Culture, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Changes in the Expression and Functional Activities of C-X-C Motif Chemokine Ligand 13 ( CXCL13 ) in Hyperplastic Prostate

doi: 10.3390/ijms24010056

Figure Lengend Snippet: The expression of CXCL13 in human serum, prostate tissue and cell lines. ( A ) Serum CXCL13 concentration in BPH patients and healthy controls. ( B , C ) The protein and mRNA expression of CXCL13 in BPH tissues and normal ones. ( D – F ) The mRNA and protein expression of CXCL13 in BPH-1 and WPMY-1 cells. Representative blots are shown. * p < 0.05, ** p < 0.01 and *** p < 0.001. ( G , H ) Immunofluorescence localization of CXCL13 in BPH tissues and normal ones. ( I , J ) Immunofluorescence localization of CXCL13 in BPH-1 and WPMY-1 cells. DAPI (blue) indicates nuclear staining and Cy3-immunofluorescence (red) indicates CXCL13 protein staining. Representative graphs are shown. All scale bars are 100 μm.

Article Snippet: Human benign prostatic enlargement epithelia cell line BPH-1 (Cat. BNCC339850) was purchased from the Procell Co., Ltd. in Wuhan, China.

Techniques: Expressing, Concentration Assay, Immunofluorescence, Staining

Journal: International Journal of Molecular Sciences

Article Title: Changes in the Expression and Functional Activities of C-X-C Motif Chemokine Ligand 13 ( CXCL13 ) in Hyperplastic Prostate

doi: 10.3390/ijms24010056

Figure Lengend Snippet: rHuCXCL13 treatment promoted proliferation of BPH-1 cells, fibrosis and inflammation of WPMY-1 cells. ( A , B ) Flow cytometry analysis of cell cycle and cell apoptosis in BPH-1 cells. ( C , D ) Statistical analysis of percentages (%) of cells at each stage and apoptotic rate (%) in BPH-1 cells. ( E ) CCK8 assay of BPH-1 cells. ( F , G ) Western blot assay of EMT-, cell-cycle- and apoptosis-related proteins in BPH-1 cells. ( H – J ) The mRNA and protein expression of markers of fibrosis and inflammation in WPMY-1 cells. * p < 0.05, ** p < 0.01 and ns means no significant difference.

Article Snippet: Human benign prostatic enlargement epithelia cell line BPH-1 (Cat. BNCC339850) was purchased from the Procell Co., Ltd. in Wuhan, China.

Techniques: Flow Cytometry, CCK-8 Assay, Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Changes in the Expression and Functional Activities of C-X-C Motif Chemokine Ligand 13 ( CXCL13 ) in Hyperplastic Prostate

doi: 10.3390/ijms24010056

Figure Lengend Snippet: Knockdown of CXCL13 inhibited proliferation, EMT and promoted apoptosis of BPH-1 cells via ERK1/2 and AKT pathway. ( A – C ) Knockdown efficiency of CXCL13 at mRNA and protein levels. ( D , F ) Statistical analysis of apoptotic rate (%) and percentages (%) of cells at each stage. ( E , G ) Flow cytometry analysis of cell apoptosis and cell cycle. ( H ) CCK8 assay of BPH-1 cells. *** means p < 0.001 between si-con and si-2, ### means p < 0.001 between si-2 and si-2+rHuCXCL13. ( I , J ) Western blot assay of cell-cycle-, cell-apoptosis- and pathway-related proteins. ( K ) Relative densitometric quantification of cell-cycle-, cell-apoptosis- and pathway-related proteins. ( L ) Western blot assay of EMT-related proteins. ( M ) Relative densitometric quantification of EMT-related proteins. * p < 0.05, ** p < 0.01, *** p < 0.001 and ns means no significant difference.

Article Snippet: Human benign prostatic enlargement epithelia cell line BPH-1 (Cat. BNCC339850) was purchased from the Procell Co., Ltd. in Wuhan, China.

Techniques: Knockdown, Flow Cytometry, CCK-8 Assay, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Changes in the Expression and Functional Activities of C-X-C Motif Chemokine Ligand 13 ( CXCL13 ) in Hyperplastic Prostate

doi: 10.3390/ijms24010056

Figure Lengend Snippet: Anti-CXCR5 (1 μg/mL) could rescue the alternations induced by rHuCXCL13 in BPH-1 cells. ( A , B ) Flow cytometry analysis of cell cycle and cell apoptosis. ( C , D ) Statistical analysis of percentages (%) of cells at each stage and apoptotic rate (%). ( E ) CCK8 assay. ( F – H ) Western blot assay of cell-cycle-, apoptosis-, EMT- and pathway-related proteins. ( I ) Relative densitometric quantification of cell-cycle-, apoptosis-, EMT- and pathway-related proteins. * p < 0.05, ** p < 0.01, *** p < 0.001 and ns means no significant difference.

Article Snippet: Human benign prostatic enlargement epithelia cell line BPH-1 (Cat. BNCC339850) was purchased from the Procell Co., Ltd. in Wuhan, China.

Techniques: Flow Cytometry, CCK-8 Assay, Western Blot