Journal: The FASEB Journal

Article Title: 12-HETER1/GPR31, a high-affinity 12( S )-hydroxyeicosatetraenoic acid receptor, is significantly up-regulated in prostate cancer and plays a critical role in prostate cancer progression

doi: 10.1096/fj.201500076

Figure Lengend Snippet: 12-HETER1 expression positively correlates with aggressiveness and progression of prostate tumors. A) Expression of 12-HETER1 in prostate carcinoma cells. Immunocytochemistry on HEK293 cells expressing 12-HETER1. a, b) Staining with 12-HETER1 antibody was optimized on HEK293 cells transfected with 12-HETER1 (a) or pcDNA (b) vector. Strong brown staining was detected in HEK293/12-HETER1 cells (a) compared to negative control (b). c, d) Immunohistochemistry to detect 12-HETER1 in 2 cases of human prostate cancer tissue. c) Benign gland (N, left corner, arrow) with rare cells showing positive staining, and neoplastic glands (T, GS-3/4, arrowhead) showing intense brown staining (arrowhead); original magnification, ×10. d) Strong staining in area (upper left, arrow) with mixed GS-3/4 tumor area, weak staining in area (lower right, arrowhead) with GS-3 tumor; original magnification, ×10. Images are representatives from 7 samples. B) Array analysis of 12-HETER1 expression. a, b) Immunohistochemistry to assay 12-HETER1 expression on 2 human prostate tissue microarrays. Expression of 12-HETER1 is significantly up-regulated in disease stage II/III human prostate tumor specimens (n = 8, P = 0.007, Student’s t test. c, d) Representative images of normal (c) and prostate tumor (d) specimens.

Article Snippet: Next we used qPCR to determine 12-HETER1 expression in 2 commercially available human prostate tissue microarrays from OriGene Technologies, HPRT101 and HPRT102 ( ).

Techniques: Expressing, Immunocytochemistry, Staining, Transfection, Plasmid Preparation, Negative Control, Immunohistochemistry

Journal: Asian Journal of Andrology

Article Title: Phosphoglycerate mutase 1 knockdown inhibits prostate cancer cell growth, migration, and invasion

doi: 10.4103/aja.aja_57_17

Figure Lengend Snippet: PGAM1 expression in prostate cancer tissues and prostate cancer cell lines. ( a ) The immunohistochemistry image of the whole TMA (scale bar = 1.5 mm). Representative immunohistochemistry images of PGAM1 protein expression from ( a ) with high intensity ( b ) in prostate cancer tissue (D7, scale bar = 0.2 mm), with intermediate intensity ( c ) in prostate cancer tissue (H7, scale bar = 0.2 mm), and with low intensity ( d ) in prostate cancer tissue (E4, scale bar = 0.2 mm). The percentage of PGAM1 positive cells in D7 is 90%, in H7 is 70%, and in E4 is 15%. Therefore, the percentage score of the case in D7 is 3 and its total protein expression score is 3 × 3 = 9. The percentage score of the case in H7 is 2 and its total protein expression score is 2 × 2 = 4. The percentage score of the case in E4 is 1 and its total protein expression score is 1 × 1 = 1. ( e ) Part of C14 from ( a ) with a magnification of 400× (scale bar = 50 μm). ( f ) Western blotting of PGAM1 protein expression in prostate cancer cell lines and normal prostate epithelial cell lines. PGAM1: phosphoglycerate mutase 1; TMA: tissue microarray.

Article Snippet: Four human PCa cell lines, PC-3, 22Rv1, DU145, and LNCap (ATCC, Manassas, VA, USA), and a human prostate epithelial cell line, RWPE1 (ATCC), were used in this study.

Techniques: Expressing, Immunohistochemistry, Western Blot, Microarray

Journal: Nature communications

Article Title: Recruitment of Mesenchymal Stem Cells Into Prostate Tumors Promotes Metastasis

doi: 10.1038/ncomms2766

Figure Lengend Snippet: ( a ) CXCR6 mRNA expression by human MSCs (P 1 and P 2 ). ( b ) Expression of CXCR6 protein by human MSCs. Controls included isotype matched controls and fibroblast-specific protein 1 (FSP1) for MSCs. Scale bars, 100μm. ( c ) CXCR6 mRNA by mMSCs. CXCR6 expression was determined in freshly isolated, non-cultured (P 0 ) or P 2 murine MSCs from CXCR6 +/+ or CXCR6 −/− mice. Human and murine osteoblasts (HOB and MC3T3-E1) were used as a negative control. ( d ) Expression of CXCR6 by murine P 2 CXCR6 +/+ or CXCR6 −/− MSCs by IHC staining. Scale bar 100μm. Data in ( a-c ) are representative of mean with standard deviation for triplicates in each of three independent experiments (Student’s t -test). ( e ) CXCL16 expression in human prostate cancer tissue microarray in . Differences noted between normal prostate ( n = 30), Gleason 4+5 ( n = 9), Gleason 6+7 ( n = 18), and Gleason 8+9 ( n = 15) (mean±s.d. Student’s t -test). Secretionof CXCL16 by human prostate cancer cell lines ( f ) and murine prostate cancer cell lines ( g ) as determined by ELISA (mean±s.d., n = 3 independent experiments, Student’s t -test). ( h ) Migration of freshly isolated, non-cultured (P 0 ) or P 2 murine MSCs from CXCR6 +/+ or CXCR6 −/− mice in response to CXCL16. The % migrated MSC was determined by hemocytometer counting (mean±s.d., n = 3 independent experiments, Student’s t -test). ( i ) CXCR6 +/+ or CXCR6 −/− mice were implanted s.c. with RM1 cells and caliper measurements of tumor growth performed over 25 days. *Significant differences between tumors grown CXCR6 +/+ and CXCR6 −/− mice (mean±s.d, for 7 animals/group, n = 3 independent experiments, P < 0.05; Student’s t -test). ( j ) % MSCs (P 0 ) present in RM1 tumors grown in CXCR6 +/+ or CXCR6 −/− mice at day 25 (mean±s.d. for 7 animals/group, n = 3 independent experiments, Student’s t -test). ( k ) SCID mice were implanted s.c. with PC3 cells mixed with MSC P0 CXCR6 +/+ or MSC P0 CXCR6 −/− cells and tumor growth was evaluated by caliper measurements over 42 days. *Significant differences between tumors grown with PC3 cells mixed with MSC P0 CXCR6 +/+ and MSC P0 CXCR6 −/− cells (mean±s.d. for n = 5 animals/group, n = 1 independent experiment, P < 0.05, Student’s t -test).

Article Snippet: The human prostate cancer cell lines PC3, LNCaP, and DU145, and murine prostate cancer cell lines RM1 and Tramp were used (American Type Culture Collection (ATCC), Rockville, MD).

Techniques: Expressing, Isolation, Cell Culture, Negative Control, Immunohistochemistry, Standard Deviation, Microarray, Enzyme-linked Immunosorbent Assay, Migration

Journal: Cell death & disease

Article Title: Lon protease 1-mediated metabolic reprogramming promotes the progression of prostate cancer.

doi: 10.1038/s41419-025-07449-8

Figure Lengend Snippet: Fig. 1 Aberrant upregulation of LONP1 is associated with poor clinical outcomes in patients with PCa. A–D LONP1 expression of patients with PCa from TCGA database. A LONP1 expression in PCa was higher than that in matched adjacent non-cancerous tissues. B LONP1 was aberrantly overexpressed in PCa tumor tissues compared to normal tissues. C Upregulated expression of LONP1 was significantly associated with Gleason Score. D Survival probability based on LONP1 expression in PCa. LONP1 high (red) group corresponds to the fourth quartile of expression, while LONP1 low (blue) group corresponds to the first quartile. E Scatterplot showing the correlation between LONP1 expression and epithelial–mesenchymal transition (EMT) score across four different PCa study cohorts, based on Bayer et al.’s work. Spearman correlation coefficient and P value are marked. F A full view of the immunohistochemistry staining of LONP1 in the PCa tissue microarray (tumor tissues, n = 97; adjacent nontumor tissues, n = 5). G–I Representative images showing immunohistochemistry staining of LONP1 in benign prostate/ PCa tissues (G), PCa tissues at different pathological stages (H), as well as PCa tissues with Gleason Score > 7 compared with those with Gleason Score ≤7 (I). The plots show the mean value for each immunoreactivity score (IRS) with statistical evaluation. Scale bar, up: 250 μm; down: 50 μm. Data were shown as mean ± SEM. **P < 0.01; ***P < 0.001.

Article Snippet: The normal human prostatic epithelial cell line (RWPE-1) and PCa cell lines (PC3, DU145, LNCaP, and LNCaP-C4-2B) were obtained from the American Type Culture Collection (ATCC) or Cell Bank of the Chinese Academy of Sciences.

Techniques: Expressing, Immunohistochemistry, Staining, Microarray

Journal: Oncotarget

Article Title: The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer

doi:

Figure Lengend Snippet: (A) Representative examples of immunohistochemical staining for CAMK2N1 in each of the clinical stages of prostate cancers as indicated. (B) Quantification of CAMK2N1 relative immunostaining intensity for each clinical stage of prostate cancers. Data is shown as mean ± SEM for N as indicated in the figure in parethesis as shown. (C) CAMK2N1 mRNA determined by quantitative PCR. Comparison was made between normal and tumorous prostate samples. (D) QT-PCR analysis of CAMK2N1 expression in prostate cell lines (RWPE1, LNCaP, DU145, PC3).

Article Snippet: Human prostate cancer DU145 and PC3 cells were obtained from ATCC and maintained in DMEM medium (Invitrogen) supplemented with 10% fetal bovine serum.

Techniques: Immunohistochemical staining, Staining, Immunostaining, Real-time Polymerase Chain Reaction, Comparison, Expressing

Journal: Oncotarget

Article Title: The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer

doi:

Figure Lengend Snippet: DU145 and PC3 cells stably overexpressed or knocked down CAMK2N1. Cells were analyzed for cell proliferation by (A-C) MTT, cell cycle by (D-E) FACS and oncogenic growth by (F-K) Colony-forming assay. Data is shown as mean ± SEM for N > 5 separate experiments.

Article Snippet: Human prostate cancer DU145 and PC3 cells were obtained from ATCC and maintained in DMEM medium (Invitrogen) supplemented with 10% fetal bovine serum.

Techniques: Stable Transfection

Journal: Oncotarget

Article Title: The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer

doi:

Figure Lengend Snippet: (A-J) Serial measurements were conducted every five days of DU145 and PC3 tumors stably expressing CAMK2N1, injected into nude mice. The data is shown as mean ± SEM for N>6 separate tumors for each group. (A, F) Images of tumors dissected out from the sacrificed mice. (B, G) The tumor size (mm3) versus days of post injection. (C, H) Reduction in tumor weight after resection at the end of experiment. (D-E, I-J) IHC staining detected the expression of CAMK2N1 in DU145 and PC3 tumor tissues of nude mice.

Article Snippet: Human prostate cancer DU145 and PC3 cells were obtained from ATCC and maintained in DMEM medium (Invitrogen) supplemented with 10% fetal bovine serum.

Techniques: Stable Transfection, Expressing, Injection, Immunohistochemistry

Journal: Oncotarget

Article Title: The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer

doi:

Figure Lengend Snippet: DU145 cells stably overexpressed CAMK2N1. These cells were analyzed for apoptosis by (A) Annexin V staining. Quantification of colony numbers and sizes were shown as mean ± SEM for N>5 separate experiments. (B) TUNEL staining was conducted to assess the effect of CAMK2N1 on cellular apoptosis in vivo . The percentage of apoptotic cells was increased in DU145 tumor tissues derived from nude mice. Quantification of TUNEL staining was shown as mean ± SEM for N=4 separate experiments. (C) IHC staining detected the expression of Bax, Bcl2, p21 and Ki67 in DU145 tumor tissue of nude mice. Overexpression of CAM2KN1 in DU145 and PC3 tumor tissue decreased Bcl-2, Ki67 protein expression and increased p21, Bax protein expression. Data for quantified IHC was shown as mean ± SEM for N = 4 tumors in each group.

Article Snippet: Human prostate cancer DU145 and PC3 cells were obtained from ATCC and maintained in DMEM medium (Invitrogen) supplemented with 10% fetal bovine serum.

Techniques: Stable Transfection, Staining, TUNEL Assay, In Vivo, Derivative Assay, Immunohistochemistry, Expressing, Over Expression

Journal: Oncotarget

Article Title: The tumor suppressive role of CAMK2N1 in castration-resistant prostate cancer

doi:

Figure Lengend Snippet: (A-B) Microarray gene expression analysis was conducted in DU145 cells stably overexpressing CAMK2N1 (>1.5-fold and p < 0.05). (A) Functional analysis of differentially expressed genes. Gene Ontology (GO) Biological Process (BP) terms were ranked by score (score > 1.3). (B) CAMK2N1 induced and repressed clusters of genes that were chosen from the pathways. (C-D) Expression levels of ErbB2, AKT, MEK1, ERK1/2, NFκβ, Bcl-2, BAX, and p21 were determined by Western blot in DU145 cells with stably overexpressing CAMK2N1. Similar changes as to those found in the microarray analysis were observed. Each figure represents three independent experiments. (E) DU145 tumors stably expressing CAMK2N1, and mRNA levels of ErbB2, Bcl-2, NF-κβ, AKT1, AR, p21, and Bax were determined by qRT-PCR. CAMK2N1 decreased ErbB2, BCL-2, NF-κβ, AKT1, AR mRNA levels and increased p21, Bax mRNA levels in DU145 tumor tissues. The data is shown as mean ± SEM for N=4 separate tumors for each group.

Article Snippet: Human prostate cancer DU145 and PC3 cells were obtained from ATCC and maintained in DMEM medium (Invitrogen) supplemented with 10% fetal bovine serum.

Techniques: Microarray, Gene Expression, Stable Transfection, Functional Assay, Expressing, Western Blot, Quantitative RT-PCR

Journal: Nature communications

Article Title: Recruitment of Mesenchymal Stem Cells Into Prostate Tumors Promotes Metastasis

doi: 10.1038/ncomms2766

Figure Lengend Snippet: ( a ) CXCR6 mRNA expression by human MSCs (P 1 and P 2 ). ( b ) Expression of CXCR6 protein by human MSCs. Controls included isotype matched controls and fibroblast-specific protein 1 (FSP1) for MSCs. Scale bars, 100μm. ( c ) CXCR6 mRNA by mMSCs. CXCR6 expression was determined in freshly isolated, non-cultured (P 0 ) or P 2 murine MSCs from CXCR6 +/+ or CXCR6 −/− mice. Human and murine osteoblasts (HOB and MC3T3-E1) were used as a negative control. ( d ) Expression of CXCR6 by murine P 2 CXCR6 +/+ or CXCR6 −/− MSCs by IHC staining. Scale bar 100μm. Data in ( a-c ) are representative of mean with standard deviation for triplicates in each of three independent experiments (Student’s t -test). ( e ) CXCL16 expression in human prostate cancer tissue microarray in . Differences noted between normal prostate ( n = 30), Gleason 4+5 ( n = 9), Gleason 6+7 ( n = 18), and Gleason 8+9 ( n = 15) (mean±s.d. Student’s t -test). Secretionof CXCL16 by human prostate cancer cell lines ( f ) and murine prostate cancer cell lines ( g ) as determined by ELISA (mean±s.d., n = 3 independent experiments, Student’s t -test). ( h ) Migration of freshly isolated, non-cultured (P 0 ) or P 2 murine MSCs from CXCR6 +/+ or CXCR6 −/− mice in response to CXCL16. The % migrated MSC was determined by hemocytometer counting (mean±s.d., n = 3 independent experiments, Student’s t -test). ( i ) CXCR6 +/+ or CXCR6 −/− mice were implanted s.c. with RM1 cells and caliper measurements of tumor growth performed over 25 days. *Significant differences between tumors grown CXCR6 +/+ and CXCR6 −/− mice (mean±s.d, for 7 animals/group, n = 3 independent experiments, P < 0.05; Student’s t -test). ( j ) % MSCs (P 0 ) present in RM1 tumors grown in CXCR6 +/+ or CXCR6 −/− mice at day 25 (mean±s.d. for 7 animals/group, n = 3 independent experiments, Student’s t -test). ( k ) SCID mice were implanted s.c. with PC3 cells mixed with MSC P0 CXCR6 +/+ or MSC P0 CXCR6 −/− cells and tumor growth was evaluated by caliper measurements over 42 days. *Significant differences between tumors grown with PC3 cells mixed with MSC P0 CXCR6 +/+ and MSC P0 CXCR6 −/− cells (mean±s.d. for n = 5 animals/group, n = 1 independent experiment, P < 0.05, Student’s t -test).

Article Snippet: The human prostate cancer cell lines PC3, LNCaP, and DU145, and murine prostate cancer cell lines RM1 and Tramp were used (American Type Culture Collection (ATCC), Rockville, MD).

Techniques: Expressing, Isolation, Cell Culture, Negative Control, Immunohistochemistry, Standard Deviation, Microarray, Enzyme-linked Immunosorbent Assay, Migration

Journal: Nature communications

Article Title: Recruitment of Mesenchymal Stem Cells Into Prostate Tumors Promotes Metastasis

doi: 10.1038/ncomms2766

Figure Lengend Snippet: ( a ) Expression of CXCL16 mRNA in the RM1 Control or RM1 shCXCL16 cells by qRT-PCR. ( b ) Secretion of CXCL16 by RM1 Control or RM1 shCXCL16 cells was determined by ELISA. ( c ) Migration of MSC CXCR6 +/+ cells was determined toward RM1 Control cells or RM1 shCXCL16 cells. Data in ( a-c ) are representative ofmean with standard deviation for triplicates in each of three independent experiments (Student’s t -test). Significance was determined using a Student’s t -test. ( d ) Experimental scheme of RM1 Control or RM1 shCXCL16 cell implantation to CXCR6 +/+ mice for examining tumor growth and MSC cell recruitment to tumors. ( e ) The tumor growth of RM1 Control or RM1 shCXCL16 cells on CXCR6 +/+ mice was evaluated by caliper measurements over 23 days. *Significant differences between tumors grown with RM1 Control and RM1 shCXCL16 cells (mean±s.d., for n = 5 animals/group, n = 2 independent experiments, P < 0.05;ANOVA). ( f ) % MSCs present in RM1 Control or RM1 shCXCL16 tumors grown in CXCR6 +/+ mice (mean±s.d., for n = 5 animals/group, n = 2 independent experiments, Student’s t -test).

Article Snippet: The human prostate cancer cell lines PC3, LNCaP, and DU145, and murine prostate cancer cell lines RM1 and Tramp were used (American Type Culture Collection (ATCC), Rockville, MD).

Techniques: Expressing, Control, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Migration, Standard Deviation

Journal: Nature communications

Article Title: Recruitment of Mesenchymal Stem Cells Into Prostate Tumors Promotes Metastasis

doi: 10.1038/ncomms2766

Figure Lengend Snippet: ( a ) Vehicle or CXCL12 treated RM1 cells, or RM1 cells co-cultured with MSCs from CXCR6 +/+ or CXCR6 −/− mice were examined by phase contrast microscopy and IHC staining for cytokeratin, E-cadherin, N-cadherin, vimentin, and α-SMA. Scale bars, 100μm. Representative images from 2 independent studies. ( b ) Western blots analysis for epithelial (E-cadherin) and mesenchymal (N-cadherin, β-catenin, snail, slug) markers. Representative images from 2 independent studies. ( c ) EMT markers in the primary tumor were examined by IHC. Colocalization of E-cadherin or N-cadherin with FSP1 was observed. More E-cadherin by prostate cancer cells (red; white arrows) was detected in close proximity to FSP1 expressing MSC cells (green; orange arrows) in tumors grown in CXCR6 −/− mice compared to tumors grown in CXCR6 +/+ mice. In contrast, more N-cadherin expressing prostate cancer cells (red; white arrows) were detected in close proximity to N-cadherin and FSP1 co-expressing CAF cells (yellow; yellow arrows) when the tumors were grown in CXCR6 +/+ mice compared to tumors grown in CXCR6 −/− mice. Blue, DAPI nuclear stain. Scale bars, 100μm. Representative images derived from n=10 mice/group). ( d ) IHC of E-cadherin or N-cadherin positive cells within benign or Gleason 4+5 prostate cancers in human prostate tissue microarrays (TMAs) (red, E-cadherin or N-cadherin, white arrows; blue, DAPI nuclear stain). Scale bars, 100μm. ( e ) Quantification of . Mean expression scores were multiplied by percent positive cells in the field. Significant differences were noted between benign ( n = 30) or Gleason 4+5 prostate ( n = 6) (mean±s.d ANOVA). ( f ) CXCR4 mRNA was determined for EMT-induced RM1 cells following CXCL12 treatment or co-culture with MSCs derived from CXCR6 +/+ or CXCR6 −/− mice (mean±s.d., n = 3 independent experiments). ( g ) More CXCR4 expressing RM1 cells (red; white arrows) were detected in close proximity to CXCR4 and FSP1 (green; orange arrows) co-expressing CAF cells (yellow; yellow arrows) when the tumors were grown in CXCR6 +/+ mice compared to tumors grown in CXCR6 −/− mice. Scale bars, 100μRepresentative images from an experiment with n=10 animals/group). ( h ) AMD3100 or anti-CXCR4 antibody prevents the development of EMT by RM1 cells following CXCL12 exposure. Scale bars, 100μm. Representative images from an experiment with n=10 animals/group.

Article Snippet: The human prostate cancer cell lines PC3, LNCaP, and DU145, and murine prostate cancer cell lines RM1 and Tramp were used (American Type Culture Collection (ATCC), Rockville, MD).

Techniques: Cell Culture, Microscopy, Immunohistochemistry, Western Blot, Expressing, Staining, Derivative Assay, Co-Culture Assay

Journal: Nature communications

Article Title: Recruitment of Mesenchymal Stem Cells Into Prostate Tumors Promotes Metastasis

doi: 10.1038/ncomms2766

Figure Lengend Snippet: (a) Migration assays were performed in Transwell® plates using 10% serum or CXCL12 as chemoattractants. Migration toward 0.5% serum was used as a negative control. (b) Blockade of CXCR4 by AMD3100 or anti-CXCR4 antibody prevents prostate cancer migration towards CXCL12 or MSCs isolated from CXCR6 +/+ , but not CXCR6 −/− animals. Data in (a,b) are representativedata from two independent studies (mean±s.d., ANOVA). Significance was determined using a Student’s t -test. RFP-labeled RM1 WT or RM1 EMT cells were incubated with vehicle or AMD3100 in vitro , and then inoculated by intra-cardiac ( i.c. ) injection into CXCR6 +/+ or CXCR6 −/− ( n = 7). Metastasis was assessed by qPCR for RFP in a number of tissues. (c,d) Number of metastatic RM1 cells following i.c. injection. *Significance between RM1 WT treated with vehicle and RM1 WT treated with AMD3100 ( P < 0.05). # Significance between RM1 WT treated with vehicle and RM1 EMT cells treated with vehicle ( P < 0.05). † Significance between RM1 EMT treated with vehicle and RM1 EMT treated with AMD3100 ( P < 0.05). Error bars represents mean±s.d., n = 2 independent experiments, P < 0.05; Student’s t -test. (e-h) RM1 cells expressing RFP were identified in the femur of CXCR6 +/+ or CXCR6 −/− mice following i.c . injection. Red arrows identify RM1 cells. White arrows identify osteoblast on the bone surface staining positive for CXCL12 expression. Scale bars, 100μm. (f,h) Quantification of Fig. 5e and Fig. 5g, respectively. The numbers of RM1 cells were quantified on the endosteal region of the 7 long bones. Endosteal regions were defined as 12 cell diameters from bone surfaces. ((Mean±s.d. ( n = 3)., ANOVA).

Article Snippet: The human prostate cancer cell lines PC3, LNCaP, and DU145, and murine prostate cancer cell lines RM1 and Tramp were used (American Type Culture Collection (ATCC), Rockville, MD).

Techniques: Migration, Negative Control, Isolation, Labeling, Incubation, In Vitro, Injection, Expressing, Staining

Journal: Scientific Reports

Article Title: Reactive oxygen species–mediated switching expression of MMP-3 in stromal fibroblasts and cancer cells during prostate cancer progression

doi: 10.1038/s41598-017-08835-9

Figure Lengend Snippet: Detection of MMP-3 and -10 expression in clinical specimens of prostate cancer patients. Representative digital images of IHC staining of MMP-3 ( a ) and MMP-10 ( b ) in paired prostate tumor and normal prostate tissues from two individual patients in sections of a human prostate tissue microarray. Left, overview image of the tissue core; right, enlargement of the indicated area. The arrow and arrowhead respectively indicate positive staining in stromal and epithelial cells. Serum samples from benign prostatic hyperplasia (BPH) patients ( n = 12) and prostate cancer patients with a Gleason score ≤ 7 (low grade, n = 8) or ≥8 (high grade, n = 40) were tested for MMP-3 through an ELISA ( c ). Statistical analysis was performed using unpaired Student’s t -tests.

Article Snippet: Human prostate cancer epithelial cell lines, LNCaP, C4-2, PC3, PC3M, and DU145, and a human bone stromal cell line HS27A (ATCC, Manassas, VA) that were used in our previous studies – were maintained in T-medium (Invitrogen, Carlsbad, CA) with 5% fetal bovine serum (FBS; Invitrogen).

Techniques: Expressing, Immunohistochemistry, Microarray, Staining, Enzyme-linked Immunosorbent Assay

Journal: Asian Journal of Andrology

Article Title: Phosphoglycerate mutase 1 knockdown inhibits prostate cancer cell growth, migration, and invasion

doi: 10.4103/aja.aja_57_17

Figure Lengend Snippet: PGAM1 expression in prostate cancer tissues and prostate cancer cell lines. ( a ) The immunohistochemistry image of the whole TMA (scale bar = 1.5 mm). Representative immunohistochemistry images of PGAM1 protein expression from ( a ) with high intensity ( b ) in prostate cancer tissue (D7, scale bar = 0.2 mm), with intermediate intensity ( c ) in prostate cancer tissue (H7, scale bar = 0.2 mm), and with low intensity ( d ) in prostate cancer tissue (E4, scale bar = 0.2 mm). The percentage of PGAM1 positive cells in D7 is 90%, in H7 is 70%, and in E4 is 15%. Therefore, the percentage score of the case in D7 is 3 and its total protein expression score is 3 × 3 = 9. The percentage score of the case in H7 is 2 and its total protein expression score is 2 × 2 = 4. The percentage score of the case in E4 is 1 and its total protein expression score is 1 × 1 = 1. ( e ) Part of C14 from ( a ) with a magnification of 400× (scale bar = 50 μm). ( f ) Western blotting of PGAM1 protein expression in prostate cancer cell lines and normal prostate epithelial cell lines. PGAM1: phosphoglycerate mutase 1; TMA: tissue microarray.

Article Snippet: A human prostate tissue microarray (TMA; PR1921) was purchased from US Biomax, Inc. (Rockville, MD, USA).

Techniques: Expressing, Immunohistochemistry, Western Blot, Microarray