prostatic epithelial cells wpmy1  (ATCC)

Journal: Experimental & Molecular Medicine

Article Title: A ESRP1/circPHGDH/miR-149/RAP1B positive feedback loop promotes the malignant behaviors and glycolysis of prostate cancer cell

doi: 10.1038/s12276-026-01646-x

Figure Lengend Snippet: a A qPCR was applied to detect circPHGDH expression in paired PCa and para-PCa tissues ( n = 51). b The circPHGDH expression was measured by qPCR in normal cells (WPMY1) and PCa cell lines (22Rv1, DU145, DuCaP, VCaP, LNCaP and C4-2). c The circPHGDH expression detected by qPCR to confirm successful transfection. d – i After circPHGDH was knocked down or overexpressed, cell proliferation ( d ) was analyzed by colony formation analysis; migration ( e ) and invasion ( f ) were estimated by Transwell assay; E-cadherin and N-cadherin protein levels ( g ) were examined using western blotting; and ECAR ( h ) and OCR ( i ) were measured using Seahorse assay to reflect glycolytic fluxes. n = 3. * P < 0.05, ** P < 0.01, *** P < 0.001, ## P < 0.01 versus the oe-nc group.

Article Snippet: Prostatic epithelial cells (WPMY1) and PCa cell lines (22Rv1, DU145, DuCaP, VCaP, LNCaP and C4-2), obtained from the American Type Culture Collection (ATCC), were cultured in Dulbecco’s modified Eagle medium (Thermo Fisher Scientific) supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin at 37 °C in a 5% CO 2 .

Techniques: Expressing, Transfection, Migration, Transwell Assay, Western Blot

Journal: Experimental & Molecular Medicine

Article Title: A ESRP1/circPHGDH/miR-149/RAP1B positive feedback loop promotes the malignant behaviors and glycolysis of prostate cancer cell

doi: 10.1038/s12276-026-01646-x

Figure Lengend Snippet: a Targeted miRNAs were chosen by a luciferase reporter analysis. b The predicted complementary sequences of circPHGDH and miR-149. c , d A dual-luciferase reporter assay ( c ) and RNA pull-down analysis ( d ) were determined for targeting relationship confirmation. e A qPCR was applied to test miR-149 expression in PCa cells after sh-circPHGDH or oe-circPHGDH transfection. f The MiR-149 expression in PCa tissues and para-PCa tissues was examined by qPCR ( n = 51). g The correlation of miR-149 and circPHGDH expression in PCa tissues was analyzed using Pearson correlation coefficient. h The MiR-149 expression in WPMY1 and PCa cell lines was tested by qPCR. i The location of circPHGDH and miR-149 in PCa cells was observed using FISH. n = 3. * P < 0.05, ** P < 0.01, *** P < 0.001, ## P < 0.01 versus the oe-nc group.

Article Snippet: Prostatic epithelial cells (WPMY1) and PCa cell lines (22Rv1, DU145, DuCaP, VCaP, LNCaP and C4-2), obtained from the American Type Culture Collection (ATCC), were cultured in Dulbecco’s modified Eagle medium (Thermo Fisher Scientific) supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin at 37 °C in a 5% CO 2 .

Techniques: Luciferase, Reporter Assay, Expressing, Transfection

Journal: Experimental & Molecular Medicine

Article Title: A ESRP1/circPHGDH/miR-149/RAP1B positive feedback loop promotes the malignant behaviors and glycolysis of prostate cancer cell

doi: 10.1038/s12276-026-01646-x

Figure Lengend Snippet: a The targets of miR-149 predicted using the miRDB, starbase and TargetScan databases are shown using Venn diagram. b The KEGG enrichment analysis of these targeted genes. c The predicted complementary sequences of RAP1B and miR-149. d , e Luciferase reporter ( d ) and RNA pull-down assays ( e ) were used to determine the targeting relationship. f , g qPCR ( f ) and western blotting ( g ) were applied to detect RAP1B expression after circPHGDH and miR-149 knockdown. h The RAP1B levels in PCa tissues and para-PCa tissues were detected by qPCR ( n = 51). i The correlation of miR-149 and RAP1B expression in PCa tissues were analyzed using Pearson correlation coefficient. j , k The RAP1B levels in WPMY1 and PCa cell lines were tested by qPCR ( j ) and western blotting ( k ). l The RAP1B-correlated genes were predicted using the LinkedOmics database. m The KEGG enrichment analysis of all RAP1B-correlated genes. n The 22Rv1 and VCap cells were transfected RAP1B overexpression plasmids, the protein levels of RAP1B, phosphorylated (p)-PI3K, PI3K, p-AKT and AKT were detected using western blotting. n = 3. * P < 0.05, ** P < 0.01, *** P < 0.001, ## P < 0.01 versus the sh-circPHGDH + nc inhibitor group.

Article Snippet: Prostatic epithelial cells (WPMY1) and PCa cell lines (22Rv1, DU145, DuCaP, VCaP, LNCaP and C4-2), obtained from the American Type Culture Collection (ATCC), were cultured in Dulbecco’s modified Eagle medium (Thermo Fisher Scientific) supplemented with 10% fetal bovine serum and 1% penicillin–streptomycin at 37 °C in a 5% CO 2 .

Techniques: Luciferase, Western Blot, Expressing, Knockdown, Transfection, Over Expression