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Updated: Jan 6, 2026
3-D Cell Culture System for Studying Invasion and Evaluating Therapeutics in Bladder Cancer
Published on: September 13, 2018
ZSTK474 suppresses bladder cancer cell migration and invasion via PI3K/Akt/Girdin pathway: an in vitro study
Zhaoxia Song1, Ying Wang1, Chunhu Yu1
1Department of Urology, Beijing Rehabilitation Hospital Affiliated to Capital Medical University, Xixiazhuang, Badachu Road, Shijingshan District, Beijing, 100144, China.
Objective:
Metastasis is the leading cause of mortality in bladder cancer (BC). This study investigates the antimetastatic potential of ZSTK474, a phosphatidylinositol 3-kinase (PI3K) inhibitor developed for solid tumor therapy. Specifically, the study evaluates the effects of ZSTK474 on tumor cell migration and invasion, two critical processes in metastasis using in vitro assays.
Methods:
Human BC cell lines, CAL29 and BIU87, were treated with ZSTK474 at concentrations of 0, 0.05, 0.25, and 1.5 µM. Cell proliferation was assessed using the BrdU incorporation assay. Migration capacity was evaluated through scratch and Transwell migration assays, while invasion potential was determined using a Matrigel-coated Transwell invasion assay. Western blotting was performed to analyze the expression levels of Girdin, phosphorylated Girdin (p-Girdin), phosphorylated Akt (p-Akt), and PI3K.
Results:
Treatment with ZSTK474 significantly inhibited BC cell proliferation, as demonstrated by the BrdU incorporation assay. Additionally, a substantial reduction in cell invasion was observed in the Transwell invasion assay (p < 0.01). Western blot analysis revealed that ZSTK474 suppressed the phosphorylation of Girdin in BC cells. Higher concentrations of ZSTK474 were associated with decreased relative expression levels of Girdin, p-Girdin, p-Akt, and PI3K. However, no significant inhibitory effect on Akt expression was detected following ZSTK474 treatment. Although total Akt protein levels remained unchanged, whereas phosphorylated Akt (p-Akt) was significantly reduced.
Conclusion:
The findings of this study suggest that ZSTK474 effectively inhibits bladder cancer cell migration and invasion by suppressing key signaling pathways involved in metastasis. Specifically, ZSTK474 reduces the phosphorylation of Girdin, an Akt substrate implicated in tumor migration and invasion. These results provide further evidence supporting the therapeutic potential of ZSTK474 in mitigating metastatic progression in bladder cancer.
Insights
ZSTK474, a PI3K inhibitor, significantly reduced bladder cancer cell proliferation, migration, and invasion in vitro. It suppressed Girdin phosphorylation, a key factor in metastasis, indicating therapeutic potential.
Area of Science:
- Oncology
- Molecular Biology
- Cancer Research
Background:
- Metastasis is the primary cause of mortality in bladder cancer (BC).
- Phosphatidylinositol 3-kinase (PI3K) signaling pathways are crucial in cancer progression.
- ZSTK474 is a developed PI3K inhibitor for solid tumor therapy.
Purpose of the Study:
- To investigate the antimetastatic potential of ZSTK474 in bladder cancer.
- To evaluate the effects of ZSTK474 on BC cell migration and invasion.
- To analyze the impact of ZSTK474 on key signaling molecules involved in metastasis.
Main Methods:
- Human BC cell lines (CAL29, BIU87) were treated with varying concentrations of ZSTK474.
- Cell proliferation was assessed using BrdU incorporation assays.
- Migration and invasion were evaluated using scratch, Transwell migration, and Matrigel-coated Transwell invasion assays.
- Western blotting analyzed Girdin, p-Girdin, p-Akt, and PI3K expression.
Main Results:
- ZSTK474 significantly inhibited BC cell proliferation and invasion (p < 0.01).
- ZSTK474 suppressed Girdin phosphorylation in BC cells.
- Higher ZSTK474 concentrations reduced Girdin, p-Girdin, p-Akt, and PI3K levels, but not total Akt.
Conclusions:
- ZSTK474 demonstrates significant antimetastatic effects in bladder cancer.
- The drug inhibits BC cell migration and invasion by suppressing Girdin phosphorylation.
- ZSTK474 shows therapeutic promise for reducing metastatic progression in bladder cancer.
