c4 2b enzr cell lines  (ATCC)

Journal: Molecular Biomedicine

Article Title: Targeting the RNA-binding motif protein 15 suppresses prostate cancer progression and hormone therapy resistance by promoting androgen receptor degradation

doi: 10.1186/s43556-026-00428-1

Figure Lengend Snippet: RBM15 silencing inhibits growth, migration, and invasion, as well as promotes apoptosis of PCa cells. a - b Representative ( a ) and statistical ( b ) analysis of RBM15 knockdown and RBM15b expression by lentivirus infection in PCa cells was evaluated using western blot (samples from 4 independent experiments). c CCK-8 experiments were employed to determine the changes in cell viability within 96 h following RBM15 knockdown. d Changes in the proliferative activity of C4-2B EnzR and 22Rv1 cells after RBM15 knockdown were examined by EdU assays ( n = 3). e Colony formation assays were used to evaluate the impact of RBM15 silencing on the proliferative capacity of cells. f Flow cytometry was adopted to examine changes in various phases of the cell cycle following RBM15 knockdown ( n = 3). g Changes in the invasion capabilities of C4-2B EnzR and 22Rv1 cells after RBM15 knockdown were assessed using transwell invasion assays (samples from 3 independent experiments). h The impact of RBM15 silencing on the apoptosis rate of PCa cells was evaluated by flow cytometry. The data are shown as mean ± SD. KD and CON represent RBM15 knockdown and the corresponding control group, respectively. SiNC denotes the negative control of siRNA. Two-tailed Student’s t test for d and One-way Anova test for b , c , f , g , h

Article Snippet: The human prostate cancer cells used in this study included the 22Rv1, LNCaP and C4-2B EnzR cell lines, which were obtained from American Type Culture Collection (Manassas, VA, USA).

Techniques: Migration, Knockdown, Expressing, Infection, Western Blot, CCK-8 Assay, Activity Assay, Flow Cytometry, Control, Negative Control, Two Tailed Test

Journal: Molecular Biomedicine

Article Title: Targeting the RNA-binding motif protein 15 suppresses prostate cancer progression and hormone therapy resistance by promoting androgen receptor degradation

doi: 10.1186/s43556-026-00428-1

Figure Lengend Snippet: RBM15 promotes resistance to enzalutamide in PCa cells. a - b The effect of different concentrations of enzalutamide on the viability of C4-2B EnzR cells with RBM15 knockdown or overexpression. c - d Difference in the effect of enzalutamide on 22RV-1 cell viability between cells with and without RBM15 knockdown or overexpression (samples from 4 independent experiments). e – f Colony formation assays evaluating the impact of knocking down ( e ) or overexpressing ( f ). g - j The effect of knocking down ( g , h ) or overexpressing ( i , j ) RBM15, combined with or without enzalutamide treatment, on the expression of AR and its downstream target PSA (samples from 4 independent experiments). Cells were treated with enzalutamide for 3 days. ns denotes no significance. Enz denotes enzalutamide. KD and OE represent RBM15 knockdown and overexpression, respectively. Scramble and vector serve as the negative controls for the knockdown and overexpression of RBM15 groups, respectively. Data are displayed as mean ± SD. Two-tailed Student’s t test

Article Snippet: The human prostate cancer cells used in this study included the 22Rv1, LNCaP and C4-2B EnzR cell lines, which were obtained from American Type Culture Collection (Manassas, VA, USA).

Techniques: Knockdown, Over Expression, Expressing, Plasmid Preparation, Two Tailed Test

Journal: Molecular Biomedicine

Article Title: Targeting the RNA-binding motif protein 15 suppresses prostate cancer progression and hormone therapy resistance by promoting androgen receptor degradation

doi: 10.1186/s43556-026-00428-1

Figure Lengend Snippet: RBM15 inhibits AR degradation through the ubiquitin–proteasome pathway. a GO enrichment analysis was performed on significantly differentially expressed genes caused by RBM15 overexpression in RNA sequencing data. b In the GSE73893 dataset, GO enrichment analyses were conducted on the RNA bound by RBM15 protein. c The impact of RBM15 knockdown on AR protein degradation under various exposure times to 20 μg/mL cycloheximide ( n = 3). d - e The impact of RBM15 overexpression on AR protein degradation under various exposure times to 20 μg/mL cycloheximide in 22Rv1 cells ( n = 3). f C4-2B EnzR cells were knocked down by shRBM15 lentivirus and then treated with 10 μM KG132 or chloroquine for 12 h. The expression of AR proteins was measured using western blotting. g Changes in AR protein expression were observed after 0, 4, 8 h of MG132 treatment in C4-2B EnzR cells, following lentiviral-mediated RBM15 knockdown ( n = 3). h - i Coimmunoprecipitation experiments revealing the effects of RBM15 knockdown ( h ) or overexpression ( i ) on the ubiquitination levels of AR proteins. Data are shown mean ± SD. CHX represents cycloheximide. CQ denotes chloroquine. One-way Anova test

Article Snippet: The human prostate cancer cells used in this study included the 22Rv1, LNCaP and C4-2B EnzR cell lines, which were obtained from American Type Culture Collection (Manassas, VA, USA).

Techniques: Ubiquitin Proteomics, Over Expression, RNA Sequencing, Knockdown, Expressing, Western Blot

Journal: Molecular Biomedicine

Article Title: Targeting the RNA-binding motif protein 15 suppresses prostate cancer progression and hormone therapy resistance by promoting androgen receptor degradation

doi: 10.1186/s43556-026-00428-1

Figure Lengend Snippet: RBM15 upregulates AR expression by decreasing K48-linked polyubiquitination levels of AR via DDB1. a Integrative analysis of the mass spectrometry of coimmunoprecipitation with anti-AR antibody, RBM15-overexpressing RNA-seq, and RBM15 RIP-seq data to identify molecules that mediate RBM15’s regulation of AR. b Coimmunoprecipitation experiments were conducted to detect the protein interaction between AR and DDB1 in 22Rv1 cells. c HA-tagged AR and Flag-tagged DDB1 plasmids were transfected into 293 T cells for 72 h, followed by immunoprecipitation experiments to assess the protein interaction between DDB1 and AR. d Images captured by confocal microscopy showing the cellular distribution of AR and DDB1 proteins in 22Rv1 cells. e Western blotting was employed to evaluate the efficiency of DDB1 knockdown in 22Rv1 cells via siRNA and DDB1 overexpression in C4-2B EnzR cells using a plasmid transfection. f The impact of silencing and overexpressing DDB1 on AR protein expression in 22Rv1 and C4-2B EnzR cells, respectively. g Alterations of AR protein expression in C4-2B EnzR cells due to DDB1 overexpression alone or in combination with MG132 treatment. h - i Immunoprecipitation assays assessed the effect of knocking down ( h ) and overexpressing ( i ) DDB1 on the ubiquitination levels of AR proteins in 22Rv1 and C4-2B EnzR cells, respectively. j Western blotting showing that RBM15 could regulate DDB1 protein expression in C4-2B EnzR and 22Rv1 cells. k Rescue assays exhibiting that DDB1 could reverse the RBM15-induced alterations of AR and AR-V7 proteins in C4-2B EnzR and 22Rv1 cells. l Immunoprecipitation assays showing that DDB1 reversed the RBM15-induced changes in ubiquitination levels of AR proteins in C4-2B EnzR and 22Rv1 cells. m Western blotting demonstrated that RBM15 regulated the K48-linked polyubiquitination levels of AR in 293 T cells, which could be reversed by DDB1. CON denotes the control group

Article Snippet: The human prostate cancer cells used in this study included the 22Rv1, LNCaP and C4-2B EnzR cell lines, which were obtained from American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing, Mass Spectrometry, RNA Sequencing, Transfection, Immunoprecipitation, Confocal Microscopy, Western Blot, Knockdown, Over Expression, Plasmid Preparation, Ubiquitin Proteomics, Control