Journal: Science signaling

Article Title: PSMA redirects cell survival signaling from the MAPK to the PI3K-AKT pathways to promote the progression of prostate cancer.

doi: 10.1126/scisignal.aag3326

Figure Lengend Snippet: Fig. 4. Manipulation of PSMA in both mouse TRAMP-C1 and human 22RV1 and PC-3 PCa cell lines mimic pathway switch. (A) CRISPR knockdown of PSMA in both the mouse TRAMP-C1 cell line (TRAMP-PSMAKO) and the human 22Rv1 cell line (22RV1-PSMAKO). Western blot analysis of both CRISPR cell lines to exam- ine changes in PDK-Ser241, IGF-1R, survivin, and pERK1/2 compared to controls (Scr). (B) TRAMP-C1 cells transiently transfected with peptides blocking the PSMA NH2-terminal cytoplasmic tail. Western blot at 24 hours for pERK1/2, cleaved PARP Asp214, and survivin at 24 hours compared to the control (Scr). (C) Western blot analysis of PC-3 cells expressing human PSMA (hPSMA) and empty vector (EV) control cell lysates to investigate changes in the previously identified signaling pathways (PSMA, IGF-1R, AKT-Ser380, AKT-Thr308, PDK1-Ser241, pERK1/2, survivin, and b-actin). All data are representative images from the mean ± SE of n = 3 for each experimental condition and three experimental replicates normalized to b-actin and presented as fold change, where WT is equal to 1. *P < 0.05, paired Student’s t test.

Article Snippet: 10 of 13 SC I ENCE S I GNAL ING | R E S EARCH ART I C L E D ow nloaded from https://w w w .science.org on January 23, 2024 CRISPR knockout of PSMA in PCa cell lines TRAMP-C1 and 22RV1 PSMA CRISPR/Cas9 constructs were created in both TRAMP-C1 and 22RV1 cell lines using the lentiCRISPRv2 plasmid #52961 (Addgene) (85, 86).

Techniques: CRISPR, Knockdown, Western Blot, Transfection, Blocking Assay, Control, Expressing, Plasmid Preparation, Protein-Protein interactions

Journal: Science signaling

Article Title: PSMA redirects cell survival signaling from the MAPK to the PI3K-AKT pathways to promote the progression of prostate cancer.

doi: 10.1126/scisignal.aag3326

Figure Lengend Snippet: Fig. 5. PSMA expression markedly alters FAK phosphorylation, and PSMA is in a complex with RACK1 and IGF-1R. (A) Western analysis for FAK-Tyr397 and FAK-Tyr925 in tumor lysates from 18-week-old WT and KO mice. (B) Western blot analysis for FAK-Tyr397 in parental WT, control scramble 22Rv1 cells (Scr), and 22Rv1-PSMAKO cells. (C) Immu- noprecipitation (IP) of PSMA in WT 22Rv1 cells using PSMA monoclonal antibody or rabbit immunoglobulin G (IgG) control and Western blot for IGF-1R (arrow indicates band). Input refers to unbound fraction. (D) Immuno- precipitation of PSMA in WT 22Rv1 cells using PSMA rabbit monoclonal antibody or rabbit IgG control and Western blot for RACK1. (E) Immunoprecipitation of RACK1 in both 22Rv1-PSMAScr and 22Rv-PSMAKO cells and Western blot analysis for b1 integrin. (F and G) Both 22Rv1-PSMAScr and 22Rv-PSMAKO cells were put either not in suspension (adherent) or in suspension (nonadherent) for 2 hours and assayed by Western blot for the direct activation of FAK- Tyr925 (F) and ERK (G) by addition of extracellular matrix (ECM) for 30 min. All data are representative images from the means ± SE of n = 3 for each experimental conditions and three experimental replicates normalized to b-actin and presented as fold change, where WT is equal to 1. *P < 0.05, paired Student’s t test.

Article Snippet: 10 of 13 SC I ENCE S I GNAL ING | R E S EARCH ART I C L E D ow nloaded from https://w w w .science.org on January 23, 2024 CRISPR knockout of PSMA in PCa cell lines TRAMP-C1 and 22RV1 PSMA CRISPR/Cas9 constructs were created in both TRAMP-C1 and 22RV1 cell lines using the lentiCRISPRv2 plasmid #52961 (Addgene) (85, 86).

Techniques: Expressing, Phospho-proteomics, Western Blot, Control, Immunoprecipitation, Suspension, Activation Assay

Journal: Journal of Food and Drug Analysis

Article Title: Biological and clinical significance of the AGE-RAGE axis in the aggressiveness and prognosis of prostate cancer

doi: 10.38212/2224-6614.3475

Figure Lengend Snippet: AGEs promote cell proliferation and decrease sensitivity to chemodrugs in PCa cells. (A) Androgen-dependent LNCaP and 22rv1, and (B) androgen-independent DU145 and PC3 cells were treated for 48 h with five types of AGEs-BSA (10–500 μg/mL) derived from different glycated agents. Cell viability was measured using the MTS assay and expressed as a relative percentage of the BSA control. GA-derived and GOA-derived AGEs increased PCa cell proliferation, whereas the other AGEs failed to enhance cell proliferation. (C) Treatment with anti-RAGE antibodies inhibited GA-derived AGEs-induced cell proliferation. PCa cells were co-treated with GA-derived AGE and anti-RAGE antibodies (10 μg/mL) for 48 h. Treatment of (D) AGEs-BSA (500 μg/mL) and (E) AGE compounds increased resistance to chemodrug toxicity in PCa cells. The cells were co-treated with AGEs and DTX (7.2 μM) for 24 h, and the cell viability was measured. GA-derived and GOA-derived AGEs, CEL, and CML increased PCa cell survival in response to the DTX challenge. (F) sRAGE counteracted CML-mediated chemoresistance and rendered PCa cells more sensitive to DTX toxicity. The cells were preincubated with sRAGE for 24 h, after which DTX was added and incubated for another 24 h. Data shown represents the means ± SD from three independent experiments with 6–8 replicates each. *P < 0.05 when compared with the BSA or vehicle control. DTX, docetaxel; anti-RAGE ab, anti-RAGE antibody; sRAGE, soluble RAGE.

Article Snippet: The human PCa cell lines 22rv1 and LNCaP (androgen-dependent), DU145 and PC3 (androgen-independent), CA-HPV-10 (prostatic adenocarcinoma of Gleason grade 4/4), and PZ-HPV-7 (epithelial cells from the peripheral zone of the normal prostate) were obtained from the Bioresource Collection and Research Center (Food Industry Research and Development Institute, Taiwan).

Techniques: Derivative Assay, MTS Assay, Control, Incubation

Journal: iScience

Article Title: Systematic fine-mapping and functional studies of prostate cancer risk variants

doi: 10.1016/j.isci.2023.106497

Figure Lengend Snippet: HOTTIP re-expression rescues defective phenotypes seen in enhancer KO 22Rv1 cells

Article Snippet: Cell lines 22Rv1、LNCaP、C4-2B cell lines were obtained from ATCC.

Techniques: Expressing

Journal: iScience

Article Title: Systematic fine-mapping and functional studies of prostate cancer risk variants

doi: 10.1016/j.isci.2023.106497

Figure Lengend Snippet: Key resources table

Article Snippet: Cell lines 22Rv1、LNCaP、C4-2B cell lines were obtained from ATCC.

Techniques: Recombinant, Reporter Assay, Plasmid Preparation, Luciferase, Software