The effect of Pokemon on bladder cancer epithelial-mesenchymal transition
Changcheng Guo1, Kai Zhu1, Wei Sun1
1Department of Urology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai 200072, People's Republic of China.
Objective:
This study aimed at detecting Pokemon expression in bladder cancer cell and investigating the relationship between Pokemon and epithelial-mesenchymal transition. Furthermore, we investigated the functions of Pokemon in the carcinogenesis and development of bladder cancer. This study was also designed to observe the inhibitory effects of siRNA expression vector on Pokemon in bladder cancer cell.
Methods:
The siRNA expression vectors which were constructed to express a short hairpin RNA against Pokemon were transfected to the bladder cancer cells T24 with a liposome. Levels of Pokemon, E-cadherin and β-catenin mRNA and protein were examined by real-time quantitative-fluorescent PCR and Western blot analysis, respectively. The effects of Pokemon silencing on epithelial-mesenchymal transition of T24 cells were evaluated with wound-healing assay.
Results:
Pokemon was strongly inhibited by siRNA treatment, especially siRNA3 treatment group, as it was reflected by Western blot and real-time PCR. The gene and protein of E-cadherin expression level showed increased markedly after Pokemon was inhibited by RNA interference. While there were no differences in the levels of gene and protein of β-catenin among five groups. The bladder cancer cell after Pokemon siRNA interference showed a significantly reduced wound-closing efficiency at 6, 12 and 24h.
Conclusions:
Our findings suggest Pokemon may inhibit the expression of E-cadherin. The low expression of E-cadherin lead to increasing the phenotype and apical-base polarity of epithelial cells. These changes of cells may result in the recurrence and progression of bladder cancer at last.
Insights
Pokemon (a protein) may drive bladder cancer progression by suppressing E-cadherin. Inhibiting Pokemon with siRNA reduced cancer cell migration, suggesting a therapeutic target for bladder cancer treatment.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Epithelial-mesenchymal transition (EMT) is crucial for cancer progression.
- The role of Pokemon in bladder cancer development and its association with EMT requires further investigation.
Purpose of the Study:
- To detect Pokemon expression in bladder cancer cells.
- To investigate the relationship between Pokemon and EMT.
- To explore Pokemon's function in bladder cancer carcinogenesis and development.
- To assess the inhibitory effects of siRNA on Pokemon in bladder cancer cells.
Main Methods:
- siRNA expression vectors targeting Pokemon were transfected into T24 bladder cancer cells.
- Pokemon, E-cadherin, and β-catenin mRNA and protein levels were quantified using real-time PCR and Western blot.
- The impact of Pokemon silencing on EMT was assessed via wound-healing assays.
Main Results:
- Pokemon expression was significantly inhibited by siRNA, particularly siRNA3.
- Inhibition of Pokemon led to a marked increase in E-cadherin expression at both gene and protein levels.
- β-catenin levels remained unchanged.
- Pokemon silencing reduced the wound-closing efficiency of bladder cancer cells.
Conclusions:
- Pokemon may inhibit E-cadherin expression, potentially promoting EMT.
- Reduced E-cadherin expression is linked to increased epithelial cell plasticity and polarity changes.
- These cellular alterations may contribute to bladder cancer recurrence and progression.
