Journal: Frontiers in Oncology

Article Title: ADGRD1 promotes bladder cancer progression and angiogenesis via the PI3K/AKT/mTOR-mediated pro-angiogenic secretome

doi: 10.3389/fonc.2026.1817872

Figure Lengend Snippet: ADGRD1 is upregulated in bladder cancer and correlates with poor prognosis. (A) GEPIA database analysis showing increased ADGRD1 expression in bladder cancer (BLCA) tissues with advancing tumor stage (Stage II–IV). (B) Kaplan–Meier survival curves from GEPIA3 demonstrating that high ADGRD1 expression is associated with reduced overall survival (OS) and progression-free interval (PFI). (C) qPCR analysis of ADGRD1 mRNA levels in non-invasive (5637, RT112, HT1376) and metastatic (UMUC1, T24, 253JBV) BLCA cell lines. (D) Western blot showing higher ADGRD1 protein expression in metastatic BLCA cells compared with non-invasive counterparts. Data are presented as mean ± SD of three independent biological replicates. *P < 0.05, **P < 0.01, ***P < 0.001 vs 5637.

Article Snippet: Human bladder cancer cell lines 5637, RT112, HT1376, and T24 (Procell, Wuhan, China), UMUC1 (Mingzhoubio, Zhejiang, China), and 253JBV (JiningBio, Shanghai, China) were maintained in RPMI-1640 medium (Gibco, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco, USA), 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Expressing, Western Blot

Journal: Frontiers in Oncology

Article Title: ADGRD1 promotes bladder cancer progression and angiogenesis via the PI3K/AKT/mTOR-mediated pro-angiogenic secretome

doi: 10.3389/fonc.2026.1817872

Figure Lengend Snippet: ADGRD1 promotes proliferation, migration, and invasion of bladder cancer cells in vitro . (A, B) Validation of ADGRD1 knockdown (sh#1, sh#2) in 253JBV and T24 cells and overexpression (OE) in 5637 cells by qPCR and Western blot. (C) CCK-8 assay showing decreased cell viability following ADGRD1 knockdown and increased viability upon ADGRD1 overexpression. (D) Colony formation assay confirming that ADGRD1 depletion inhibits, while overexpression enhances, clonogenic growth. (E) Wound-healing assay showing impaired migration in ADGRD1-silenced cells and accelerated wound closure in ADGRD1-overexpressing cells. (F) Transwell migration assay and (G) Transwell invasion assay demonstrating consistent results. Data are presented as mean ± SD of three independent biological replicates. ***P < 0.001.

Article Snippet: Human bladder cancer cell lines 5637, RT112, HT1376, and T24 (Procell, Wuhan, China), UMUC1 (Mingzhoubio, Zhejiang, China), and 253JBV (JiningBio, Shanghai, China) were maintained in RPMI-1640 medium (Gibco, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco, USA), 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Migration, In Vitro, Biomarker Discovery, Knockdown, Over Expression, Western Blot, CCK-8 Assay, Colony Assay, Wound Healing Assay, Transwell Migration Assay, Transwell Invasion Assay

Journal: Frontiers in Oncology

Article Title: ADGRD1 promotes bladder cancer progression and angiogenesis via the PI3K/AKT/mTOR-mediated pro-angiogenic secretome

doi: 10.3389/fonc.2026.1817872

Figure Lengend Snippet: ADGRD1 activates the PI3K/AKT/mTOR signaling pathway. (A) Western blot showing reduced phosphorylation of PI3K, AKT, and mTOR in ADGRD1-silenced 253JBV and T24 cells, and elevated phosphorylation in ADGRD1-overexpressing 5637 cells. (B) Immunoblotting of xenograft tumor tissues confirming consistent regulation of PI3K/AKT/mTOR phosphorylation in vivo . β-actin served as the loading control. Data are presented as mean ± SD. N = 3 independent biological replicates or 3 mice per group. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Human bladder cancer cell lines 5637, RT112, HT1376, and T24 (Procell, Wuhan, China), UMUC1 (Mingzhoubio, Zhejiang, China), and 253JBV (JiningBio, Shanghai, China) were maintained in RPMI-1640 medium (Gibco, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco, USA), 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: Western Blot, Phospho-proteomics, In Vivo, Control

Journal: Frontiers in Oncology

Article Title: ADGRD1 promotes bladder cancer progression and angiogenesis via the PI3K/AKT/mTOR-mediated pro-angiogenic secretome

doi: 10.3389/fonc.2026.1817872

Figure Lengend Snippet: Pharmacological modulation of PI3K/AKT/mTOR signaling reverses ADGRD1-mediated malignant phenotypes.ADGRD1-silenced cells (253JBV, T24) were treated with SC79 (AKT activator), while ADGRD1-overexpressing 5637 cells were treated with LY294002 (PI3K inhibitor). (A) CCK-8 assay showing that SC79 rescued, and LY294002 abolished, ADGRD1-dependent changes in cell viability. (B) Colony formation, (C) wound-healing, (D) Transwell migration, (E) Transwell invasion, and (F) HUVEC tube formation assays demonstrating consistent reversal of ADGRD1 effects upon pathway modulation. Scale bars, 100 µm. Data are presented as mean ± SD of three independent biological replicates. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Human bladder cancer cell lines 5637, RT112, HT1376, and T24 (Procell, Wuhan, China), UMUC1 (Mingzhoubio, Zhejiang, China), and 253JBV (JiningBio, Shanghai, China) were maintained in RPMI-1640 medium (Gibco, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco, USA), 100 U/mL penicillin, and 100 μg/mL streptomycin at 37 °C in a humidified incubator with 5% CO 2 .

Techniques: CCK-8 Assay, Migration