Journal: Scientific Reports

Article Title: Dynamic interplay between sortilin and syndecan-1 contributes to prostate cancer progression

doi: 10.1038/s41598-023-40347-7

Figure Lengend Snippet: Sortilin (SORT) interacts with progranulin (PRGN) and LPL in PCa cells and PRGN is secreted by aggressive PC3 cells. ( a ) Representative confocal images showing co-labelling of cells with anti-PRGN (red) and anti-SORT (green) antibodies. Scale bars; 10 µm. ( b ) Quantification of colocalisation between PRGN and SORT. ( c ) Amount of PRGN in conditioned media (CM) and corresponding cell lysates with quantification of band densities and ratio of PRGN in CM to PRGN in cell lysates. ( d ) Endogenous SORT was immunoprecipitated (IP) with anti-SORT antibodies and PRGN was detected by Western blotting. Input; cell lysate. ( e ) Representative confocal images showing co-labelling of cells with anti-LPL (red) and anti-SORT (green) antibodies. Scale bars; 10 µm. ( f ) Quantification of colocalisation between LPL and SORT. ( g ) Amount of LPL in CM and corresponding cell lysates with quantification of band densities in cell lysates. In ( c ) and ( g ) PNT2 bands from the same membrane were shifted towards LNCaP and PC3 bands. Western blotting signal was normalised using total protein staining. ( h ) Endogenous SORT was immunoprecipitated (IP) with anti-SORT antibodies and LPL was detected by Western blotting. Input; cell lysate. Data are presented as mean ± SD, in ( b , f ) n = 10 representative ROIs, in ( c , g ) n = 3 (independent experiments), one-way ANOVA.

Article Snippet: Human prostate cell PNT2 (ECACC 95012613), LNCaP (clone FCG, ECACC 89110211) and PC3 (ECACC 90112714) were obtained from the European Collection of Authenticated Cell Cultures (ECACC) from CellBank Australia (Children's Medical Research Institute, Westmead, NSW, Australia).

Techniques: Immunoprecipitation, Western Blot, Membrane, Staining

Journal: Cancer Biology & Therapy

Article Title: CPT1A mediates the succinylation of SP5 which activates transcription of PDPK1 to promote the viability and glycolysis of prostate cancer cells

doi: 10.1080/15384047.2024.2329372

Figure Lengend Snippet: CPT1A is highly expressed in PCa tissues and cell lines. (a) GEPIA was used to analyze the dysregulated expression of CPT1A in PCa. PRAD: prostate adenocarcinoma. T: tumor and N: normal. (b) The expression of CPT1A in PCa tissues and the normal tissues was detected by qPCR. (c) The expression of CPT1A in PCa cell lines and the prostate epithelial cell line (PNT2) was detected by qPCR. * p <.05 in (A). ** p < .01, *** p < .001 vs PNT2 group.

Article Snippet: HEK-293T cells, normal human epithelial prostate PNT2 cells and the PCa cell lines including DU145, LNCap, 22Rv1 and VCaP were purchased from ATCC (USA) and were maintained in Dulbecco’s modified eagle medium containing 10% fetal bovine serum and 1% penicillin/streptomycin (all from Thermo Fisher Scientific, USA).

Techniques: Expressing

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: Schistosoma Egg Antigen Induces Oncogenic Alterations in Human Prostate Cells

doi: 10.1155/2018/4675380

Figure Lengend Snippet: schSEA depletes GSH levels in prostate (PNT2) cells. Statistical significance by Student's t -test; ∗ p ≤ 0.05 (serum-fed vs. serum-starved experiments) and # p ≤ 0.05 (non-schSEA treatment vs. schSEA-treated cells/NaNO 2 -treated cells). Results are representative of three independent experiments.

Article Snippet: Normal human prostate (PNT2) cells obtained from RIKEN BioResource Centre Cell Bank (Japan) were maintained in RPMI-1640 culture medium enriched with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin and incubated at 37°C in the presence of 5% CO 2.

Techniques:

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: Schistosoma Egg Antigen Induces Oncogenic Alterations in Human Prostate Cells

doi: 10.1155/2018/4675380

Figure Lengend Snippet: Effect of SEA treatment on proliferation of PNT2 cells.

Article Snippet: Normal human prostate (PNT2) cells obtained from RIKEN BioResource Centre Cell Bank (Japan) were maintained in RPMI-1640 culture medium enriched with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin and incubated at 37°C in the presence of 5% CO 2.

Techniques:

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: Schistosoma Egg Antigen Induces Oncogenic Alterations in Human Prostate Cells

doi: 10.1155/2018/4675380

Figure Lengend Snippet: schSEA induces proliferation of prostate (PNT2). Statistical significance by Student's t -test: ∗ p ≤ 0.05 (non-schSEA treatment vs. 12.5 μ g/ml schSEA-treated cells). Results are representative of three independent experiments.

Article Snippet: Normal human prostate (PNT2) cells obtained from RIKEN BioResource Centre Cell Bank (Japan) were maintained in RPMI-1640 culture medium enriched with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin and incubated at 37°C in the presence of 5% CO 2.

Techniques:

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: Schistosoma Egg Antigen Induces Oncogenic Alterations in Human Prostate Cells

doi: 10.1155/2018/4675380

Figure Lengend Snippet: schSEA diminishes apoptosis in PNT2 cells. (a) (A) Negative control, (B) 6.25 μ g/ml schSEA, (C) 12.5 μ g/ml schSEA, and (D) serum-fed (positive control). Lower left quadrant: viable cells; lower right quadrant: early apoptotic cells; upper left quadrant: nuclear debris; upper right quadrant: late apoptotic cells. Data was acquired by Guava EasyCyte™ flow cytometer. (b) Comparison of apoptosis and cell viability among various experimental groups. Statistical significance by Student's t -test: ∗ p ≤ 0.05 (serum-fed vs. serum-starved/schSEA-treated groups). Results are representative of three independent experiments.

Article Snippet: Normal human prostate (PNT2) cells obtained from RIKEN BioResource Centre Cell Bank (Japan) were maintained in RPMI-1640 culture medium enriched with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin and incubated at 37°C in the presence of 5% CO 2.

Techniques: Negative Control, Positive Control, Flow Cytometry, Comparison

Journal: Analytical Cellular Pathology (Amsterdam)

Article Title: Schistosoma Egg Antigen Induces Oncogenic Alterations in Human Prostate Cells

doi: 10.1155/2018/4675380

Figure Lengend Snippet: schSEA diminishes nuclei condensation in PNT2 cells: (a) negative control, (b) 6.25 μ g/ml schSEA, (c) 12.5 μ g/ml schSEA, and (d) serum-fed (positive control). Condensed and morphologically altered nuclei indicated by arrows. Cells were stained with Hoechst 33258 dye and nuclei visualized with fluorescence microscope (magnification 40x) (Olympus DP 72, Japan).

Article Snippet: Normal human prostate (PNT2) cells obtained from RIKEN BioResource Centre Cell Bank (Japan) were maintained in RPMI-1640 culture medium enriched with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin and incubated at 37°C in the presence of 5% CO 2.

Techniques: Negative Control, Positive Control, Staining, Fluorescence, Microscopy

Journal: Journal of Extracellular Vesicles

Article Title: Extracellular Vesicles From Prostate Cancer‐Corrupted Osteoclasts Drive a Chain Reaction of Inflammatory Osteolysis and Tumour Progression at the Bone Metastatic Site

doi: 10.1002/jev2.70091

Figure Lengend Snippet: Metastatic PCa cells promote OC activity and induce pathological phenotype. (a) Representative IVIS image showing PCa‐transplanted xenograft mice ( N = 3) on Day 35. Tumours in the hind limbs were evaluated based on the photon radiance of cancer cell bioluminescence. (b) Representative images of HE‐, osteoclastic and osteoblastic marker TRAP/ALP‐ and cancer‐specific marker LAT1 stained sections of bone metastatic sites in the xenografted mouse which was shown in (a). All images were obtained with an all‐in‐one fluorescence microscope using a 4x objective for low‐, 10x objective for middle‐, and 40x objective for high‐magnification. Black arrowheads indicate the tumour metastatic site, and white arrowheads indicate OCs. The metastatic tumour developed in the femur. Scale bar indicates 300 µm. (c) Schematic images of the transwell co‐culture system for obtaining cancer‐associated OCs (CAOCs) and normal osteoclasts (NOCs). OC precursor RAW 264.7 cells were grown into mature OCs in the lower compartment. Mature OCs with osteolytic PC3M cells (OCP cells) and mature OCs with osteoblastic C4‐2B cells (OCC cells) were used as CAOCs. Mature OCs with blank transwells (OC cells) and mature OCs with PNT2 (OCN cells) were prepared as control NOCs. (d) Schematic images of transwell co‐culture for observing CAOCs and NOCs. (e) Representative TRAP staining images of each OC. Bars represent 100 µm. ( f) Representative TRAP staining images of each OCs treated with denosumab at the concentration of 10 µg/mL. Bars represent 100 µm.

Article Snippet: PC3M, C4‐2B, and the immortalized normal human prostatic epithelial cell line PNT2 (DS Pharma Biomedical Co., Ltd, Osaka, Japan) were cultured in RPMI 1640 medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10%‐inactivated foetal bovine serum (FBS, Gibco, Thermo Fisher Scientific) and 1% antibiotic‐antimycotic (AA) solution (Gibco, Thermo Fisher Scientific) at 37 °C in atmosphere of 95% air and 5% CO 2 .

Techniques: Activity Assay, Marker, Staining, Fluorescence, Microscopy, Co-Culture Assay, Control, Concentration Assay

Journal: Journal of Extracellular Vesicles

Article Title: Extracellular Vesicles From Prostate Cancer‐Corrupted Osteoclasts Drive a Chain Reaction of Inflammatory Osteolysis and Tumour Progression at the Bone Metastatic Site

doi: 10.1002/jev2.70091

Figure Lengend Snippet: Inflammatory‐related signalling pathways are enhanced in CAOCs. (a) Schematic protocol of co‐culture and gene expression analysis of CAOCs. (b) Heat map showing differentially expressed genes (CAOC vs. OCN, change >2.0‐fold and p < 0.05) in RAW 264.7, co‐cultured with PCa cells ( n = 2) in the presence of RANKL. (c) The number of differentially expressed genes in mature OCs co‐cultured with PC3M cells (OCP cells) or C4‐2B cells (OCC cells) compared to that in OCN cells co‐cultured with PNT2 cells (OCN cells). (d) PCA of gene expression of each type of OCs. (e) Pathway analysis of selected genes that were significantly upregulated in CAOCs compared to OCN cells. (f) GSEA of CAOC versus NOC, highlighting pro‐inflammatory phenotypes. NES: normalized enrichment score. p value was calculated using GSEA. (g) Expression levels of Il‐1b , Casp1 , Il‐6 and Tnf in gene sets of inflammation‐related genes. OC co‐cultured with a blank insert (OC cells) or PNT2 (OCN cells), and OC co‐cultured with PC3M (OCP cells) or C4‐2B (OCC cells) are presented. The blue, green, yellow and red dots represent the OC, OCN, OCP and OCC cell data, respectively.

Article Snippet: PC3M, C4‐2B, and the immortalized normal human prostatic epithelial cell line PNT2 (DS Pharma Biomedical Co., Ltd, Osaka, Japan) were cultured in RPMI 1640 medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10%‐inactivated foetal bovine serum (FBS, Gibco, Thermo Fisher Scientific) and 1% antibiotic‐antimycotic (AA) solution (Gibco, Thermo Fisher Scientific) at 37 °C in atmosphere of 95% air and 5% CO 2 .

Techniques: Co-Culture Assay, Gene Expression, Cell Culture, Expressing