Journal: Translational Oncology

Article Title: piR-43452 suppresses bladder cancer progression and enhances gemcitabine sensitivity via GTSF1/PIWIL4-mediated LRP1 mRNA destabilization

doi: 10.1016/j.tranon.2025.102626

Figure Lengend Snippet: piR-43452 regulates LRP1 expression . A. Volcano plot of differentially expressed genes in UM-UC-3 cells after piR-43452 overexpression (RNA-seq; n = 3; cutoff: P < 0.05, |log2FC| > 0).B-C. LRP1 mRNA (B) and protein (C) expression changes in UM-UC-3 and J82 cells transfected with piR-43452 mimics.D. Luciferase reporter assay comparing wildtype (WT) and mutant (MUT) LRP1 3′UTR activity in HEK-293T cells co-transfected with piR-43452 mimics. Firefly luciferase activity was normalized to Renilla.E. RIP-qPCR showing enhanced association of PIWIL4 with piR-43452 and LRP1 mRNA compared to IgG control.F. Representative IHC images of LRP1 expression in xenograft tumors derived from piR-43452-overexpressing cells.

Article Snippet: Membranes were incubated overnight at 4 °C using antibodies against N-cadherin, MMP9, MMP2, caspase-3, PARP1 (Proteintech); LRP1 (Diag Bio); HA/Flag/β-actin (Cell Signaling Technology).

Techniques: Expressing, Over Expression, RNA Sequencing, Transfection, Luciferase, Reporter Assay, Mutagenesis, Activity Assay, Control, Derivative Assay

Journal: Translational Oncology

Article Title: piR-43452 suppresses bladder cancer progression and enhances gemcitabine sensitivity via GTSF1/PIWIL4-mediated LRP1 mRNA destabilization

doi: 10.1016/j.tranon.2025.102626

Figure Lengend Snippet: The functional GTSF1/PIWIL4 complex mediates piR-43452-induced suppression of LRP1 . A. Validation of GTSF1 overexpression in UM-UC-3 and J82 cells. B. Efficiency of GTSF1 knockdown in UM-UC-3 and J82 cells, and the corresponding expression level of PIWIL4.C. RNA immunoprecipitation using an anti-HA antibody in 3HA-GTSF1-overexpressing HEK-293T cells, followed by qPCR, shows enhanced enrichment of both piR-43452 and LRP1 mRNA in the HA immunoprecipitates compared to the IgG control. D. LRP1 expression changes in UM-UC-3 and J82 cells transfected with piR-43452 mimics and GTSF1-overexpressing plasmid.E. LRP1 expression changes in UM-UC-3 and J82 cells transfected with piR-43452 mimics and siGTSF1. F. Interaction of PIWIL4 protein and GTSF1 protein detected with Co-IP. G. Efficiency of PIWIL4 knockdown in UM-UC-3 and J82 cells, and the corresponding expression level of GTSF1.H. LRP1 expression changes in UM-UC-3 and J82 cells transfected with piR-43452 mimics and siPIWIL4. (mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: Membranes were incubated overnight at 4 °C using antibodies against N-cadherin, MMP9, MMP2, caspase-3, PARP1 (Proteintech); LRP1 (Diag Bio); HA/Flag/β-actin (Cell Signaling Technology).

Techniques: Functional Assay, Biomarker Discovery, Over Expression, Knockdown, Expressing, RNA Immunoprecipitation, Control, Transfection, Plasmid Preparation, Co-Immunoprecipitation Assay

Journal: Translational Oncology

Article Title: piR-43452 suppresses bladder cancer progression and enhances gemcitabine sensitivity via GTSF1/PIWIL4-mediated LRP1 mRNA destabilization

doi: 10.1016/j.tranon.2025.102626

Figure Lengend Snippet: LRP1 is upregulated in BCa and promotes malignant phenotypes . A. Western blot analysis of LRP1 protein levels in paired BCa and adjacent normal tissues (FAHZU cohort). B. IHC staining of LRP1 in BCa tissues and matched normal urothelium. C. Kaplan-Meier analysis of overall survival in BCa patients stratified by LRP1 expression. D-E. Proliferation of UM-UC-3 and J82 cells after LRP1 knockdown as assessed by CCK-8 (D) and colony formation (E) assays. F. Transwell migration assays demonstrating reduced metastatic potential in LRP1-knockdown cells. G-H. Western blot analysis of EMT-related (G) and apoptosis-related (H) protein expression changes following LRP1 knockdown. I. Apoptosis rates in LRP1-knockdown UM-UC-3 and J82 cells. (mean ± SEM; * P < 0.05, ** P < 0.01, *** P < 0.001).

Article Snippet: Membranes were incubated overnight at 4 °C using antibodies against N-cadherin, MMP9, MMP2, caspase-3, PARP1 (Proteintech); LRP1 (Diag Bio); HA/Flag/β-actin (Cell Signaling Technology).

Techniques: Western Blot, Immunohistochemistry, Expressing, Knockdown, CCK-8 Assay, Migration

Journal: Translational Oncology

Article Title: piR-43452 suppresses bladder cancer progression and enhances gemcitabine sensitivity via GTSF1/PIWIL4-mediated LRP1 mRNA destabilization

doi: 10.1016/j.tranon.2025.102626

Figure Lengend Snippet: piR-43452 inhibits BCa malignant phenotypes through LRP1 . A. Western blot showing rescue of LRP1 expression by ectopic LRP1 overexpression in piR-43452 mimic-transfected cells.B-D. Functional rescue experiments assessing proliferation (B: CCK-8, C: colony formation) and migration (D: Transwell) in BCa cells with combined piR-43452 overexpression and LRP1 reconstitution. (mean ± SEM; *** P < 0.001).

Article Snippet: Membranes were incubated overnight at 4 °C using antibodies against N-cadherin, MMP9, MMP2, caspase-3, PARP1 (Proteintech); LRP1 (Diag Bio); HA/Flag/β-actin (Cell Signaling Technology).

Techniques: Western Blot, Expressing, Over Expression, Transfection, Functional Assay, CCK-8 Assay, Migration

Journal: Translational Oncology

Article Title: piR-43452 suppresses bladder cancer progression and enhances gemcitabine sensitivity via GTSF1/PIWIL4-mediated LRP1 mRNA destabilization

doi: 10.1016/j.tranon.2025.102626

Figure Lengend Snippet: piR-43452 and LRP1 modulate gemcitabine sensitivity . A. IC 50 values for gemcitabine in J82 cells with piR-43452 overexpression. B. IC50 values for gemcitabine in J82 cells with piR-43452 knockdown.C-D. Cell viability (B: CCK-8) and colony formation (C) in piR-43452-overexpressing J82 cells treated with gemcitabine.E. Apoptosis rates in J82 cells with piR-43452 overexpression ± gemcitabine treatment. F. IC 50 values for gemcitabine in LRP1-knockdown J82 cells. G-H. Cell viability (F: CCK-8) and colony formation (G) in LRP1-knockdown J82 cells treated with gemcitabine. I. Apoptosis rates in LRP1-knockdown J82 cells ± gemcitabine treatment. (mean ± SEM; ** P < 0.01, *** P < 0.001).

Article Snippet: Membranes were incubated overnight at 4 °C using antibodies against N-cadherin, MMP9, MMP2, caspase-3, PARP1 (Proteintech); LRP1 (Diag Bio); HA/Flag/β-actin (Cell Signaling Technology).

Techniques: Over Expression, Knockdown, CCK-8 Assay