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Published on: October 30, 2013
Effect of CTP-mediated PTEN on 5637 bladder cancer cells and the underlying molecular mechanism
Bei Yu1, Yuan Huang2, Yue Yang1
1Urological Department, The Affiliated Hospital of Chengdu University, Chengdu, Sichuan, China.
Objective:
The aim of the present study was to explore the effect of cytoplasmic transduction peptide (CTP)-phosphatase and tensin homolog (PTEN) on the proliferation, cell cycle, apoptosis, migration and invasion of bladder cancer cells and the underlying molecular mechanism.
Methods:
A eukaryotic expression vector, pTT5-CTP-PTEN, was constructed. The constructed vector was transfected into HEK 293-6E cells to express a fusion protein, CTP-PTEN. The fusion protein was purified. 5637 bladder cancer cells were cocultured with purified CTP-PTEN fusion protein. Target gene expression, protein expression, cell proliferation, cell cycle, apoptosis, cell invasion and cell migration were examined by reverse transcription polymerase chain reaction (RT-PCR), western blot, MTT assay, flow cytometry, Transwell assay, and cell scratch assay, respectively.
Results:
Both PTEN and CTP-PTEN fusion protein inhibited the proliferation, cell cycle, invasion and migration of bladder cancer cells and promoted the apoptosis of bladder cancer cells. The effect of CTP-PTEN was more significant.
Conclusions:
The fused expression of CTP and PTEN significantly increased the penetrability of the tumor suppressor gene PTEN into cancer cells. The CTP-PTEN fusion protein exhibited a significant carcinostatic effect on 5637 bladder cancer cells.
Insights
The novel CTP-PTEN fusion protein significantly inhibits bladder cancer cell proliferation, invasion, and migration while promoting apoptosis. This enhanced delivery of PTEN offers a promising therapeutic strategy for bladder cancer.
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Bladder cancer remains a significant health concern with limited effective therapeutic options.
- The tumor suppressor gene PTEN plays a crucial role in regulating cell growth and survival.
- Efficient delivery of PTEN into cancer cells is a challenge for therapeutic applications.
Purpose of the Study:
- To investigate the effects of a novel cytoplasmic transduction peptide (CTP)-PTEN fusion protein on bladder cancer cell behavior.
- To explore the underlying molecular mechanisms of CTP-PTEN's action.
- To evaluate the therapeutic potential of CTP-PTEN in bladder cancer.
Main Methods:
- Construction and purification of a CTP-PTEN fusion protein.
- Co-culture of 5637 bladder cancer cells with CTP-PTEN.
- Assessment of proliferation (MTT assay), cell cycle (flow cytometry), apoptosis (flow cytometry), invasion (Transwell assay), and migration (cell scratch assay).
- Analysis of gene and protein expression (RT-PCR, Western blot).
Main Results:
- CTP-PTEN fusion protein significantly inhibited bladder cancer cell proliferation, cell cycle progression, invasion, and migration.
- CTP-PTEN treatment markedly promoted apoptosis in bladder cancer cells.
- The inhibitory and pro-apoptotic effects of CTP-PTEN were more pronounced than PTEN alone.
Conclusions:
- The CTP tag enhances the cellular penetrability of PTEN, a crucial tumor suppressor.
- CTP-PTEN exhibits a significant carcinostatic effect on 5637 bladder cancer cells.
- CTP-PTEN represents a promising therapeutic agent for bladder cancer treatment.
