Gallic Acid Inhibits Bladder Cancer T24 Cell Progression Through Mitochondrial Dysfunction and PI3K/Akt/NF-κB
Maolin Zeng1,2, Yang Su3,4,5, Kuangyu Li6,7
1Department of Pharmacy, Renmin Hospital of Wuhan University, Wuhan, China.
Abstract:
Gallic acid (GA), a hydrolyzable tannin, has a wide range of pharmacological activities. This study revealed that, GA significantly inhibited T24 cells viability in a concentration- and time- dependent manner. The IC50 of GA stimulating T24 cells for 24, 48, and 72 h were 21.73, 18.62, and 11.59 µg/ml respectively, and the inhibition rate was significantly higher than the positive control drug selected for CCK-8 assay. Meanwhile, after GA treatment, the morphology of T24 cells were changed significantly. Moreover, GA significantly inhibited T24 cells proliferation and blocked T24 cells cycle in S phase (p < 0.001). GA induced T24 cells apoptosis (p < 0.001), accompanied by reactive oxygen species (ROS) accumulation and mitochondrial membrane potential (MMP) depolarization. Western blotting analysis showed that GA significantly increased Cleaved caspase-3, Bax, P53, and Cytochrome C (Cyt-c) proteins expression, and decreased Bcl-2, P-PI3K, P-Akt, P-IκBα, P-IKKα, and P-NF-κB p65 proteins expression in T24 cells (p < 0.05). Real-Time PCR results verified that GA significantly promoted Caspase-3, Bax, P53, and Cyt-c genes expression, and inhibited Bcl-2, PI3K, Akt, and NF-κB p65 genes expression (p < 0.001). However, on the basis of GA (IC50) stimulation, NAC (an oxidative stress inhibitor) pretreatment reversed the apoptotic rate of T24 cells and the expression of Bax, Cleaved caspase-3, P53, Bcl-2 proteins, and the MMP level in T24 cells, as well as the expression of Cyt-c protein in T24 cells mitochondria and cytoplasm. In addition, GA significantly suppressed T24 cells migration and invasion ability with VEGF protein inhibition (p < 0.001). Briefly, GA can inhibit T24 cells proliferation, metastasis and promote apoptosis, and the pro-apoptotic activity is closely associated with mitochondrial dysfunction and PI3K/Akt/NF-κB signaling suppression. Our study will help in finding a safe and effective treatment for bladder cancer.
Insights
Gallic acid (GA) effectively inhibits bladder cancer (T24) cell proliferation, migration, and invasion. This natural compound promotes apoptosis through mitochondrial dysfunction and PI3K/Akt/NF-κB signaling pathway suppression.
Area of Science:
- Pharmacology and Toxicology
- Cancer Biology
- Natural Products Chemistry
Background:
- Gallic acid (GA), a hydrolyzable tannin, exhibits diverse pharmacological activities.
- Bladder cancer remains a significant health concern, necessitating novel therapeutic strategies.
- Understanding the molecular mechanisms of natural compounds like GA in cancer is crucial.
Purpose of the Study:
- To investigate the anti-cancer effects of Gallic acid (GA) on T24 bladder cancer cells.
- To elucidate the underlying molecular mechanisms, including apoptosis induction and signaling pathway modulation.
- To assess GA's impact on cell viability, proliferation, cell cycle, migration, and invasion.
Main Methods:
- Cell viability was assessed using CCK-8 assays.
- Cell cycle, apoptosis, reactive oxygen species (ROS), and mitochondrial membrane potential (MMP) were analyzed.
- Western blotting and Real-Time PCR were employed to examine protein and gene expression, including key apoptosis and signaling pathway markers.
Main Results:
- GA significantly inhibited T24 cell viability, proliferation, migration, and invasion in a dose- and time-dependent manner.
- GA induced apoptosis, cell cycle arrest in S phase, ROS accumulation, and MMP depolarization.
- GA modulated the expression of apoptosis-related proteins (e.g., increased Bax, Cleaved caspase-3; decreased Bcl-2) and suppressed the PI3K/Akt/NF-κB signaling pathway.
Conclusions:
- Gallic acid demonstrates potent anti-cancer properties against T24 bladder cancer cells.
- GA's pro-apoptotic activity is linked to mitochondrial dysfunction and the suppression of the PI3K/Akt/NF-κB pathway.
- GA represents a potential therapeutic agent for bladder cancer treatment.
