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Published on: October 30, 2013
Cytotoxicity and apoptosis by survivin small interfering RNA in bladder cancer cells
Ja H Ku1, Soo Y Seo, Cheol Kwak
1Department of Urology, Seoul National University Hospital, Seoul, Korea.
Objective:
To investigate the effects of survivin gene RNA interference on cell growth and the cell cycle in the human bladder cancer cell line T24.
Materials And Methods:
A small interfering RNA (siRNA) targeting survivin was transfected into T24 cells using a liposome approach. Reverse transcription-polymerase chain reaction and Western blot analysis were used to examine survivin gene expression in T24 cells. Cells densities were determined by haematocytometer counts and flow cytometry was used for cell cycle analysis. Caspase-3 activity was quantified.
Results:
After treatment with survivin siRNA, the survivin gene expression in T24 cells was almost completely absent. The survivin siRNA treatment caused a profound decrease in survivin protein, which was correlated with a decrease in cell growth, G2/M arrest, and an increase in the fraction of cells undergoing apoptosis. The inhibition of survivin expression increased caspase-3 activity in T24 cells, which led to apoptosis.
Conclusions:
RNA interference can efficiently suppress survivin expression in T24 cells. Targeting survivin by siRNA may be a promising approach to block proliferation of bladder cancer cells and may provide a suitable adjuvant therapy for treatment of bladder cancer.
Insights
RNA interference effectively suppressed survivin gene expression in T24 bladder cancer cells, inhibiting growth and promoting apoptosis. This suggests survivin targeting is a promising strategy for bladder cancer treatment.
Area of Science:
- Molecular Biology
- Oncology
- Biochemistry
Background:
- Survivin is a key protein involved in cell division and survival, often overexpressed in various cancers, including bladder cancer.
- Targeting survivin presents a potential therapeutic strategy to inhibit cancer cell proliferation.
Purpose of the Study:
- To investigate the efficacy of survivin gene RNA interference (siRNA) in suppressing survivin expression.
- To evaluate the impact of survivin suppression on cell growth, cell cycle progression, and apoptosis in the T24 human bladder cancer cell line.
Main Methods:
- Transfection of T24 cells with survivin-targeting siRNA using a liposome-based delivery system.
- Assessment of survivin gene and protein expression via RT-PCR and Western blot.
- Analysis of cell proliferation, cell cycle distribution (flow cytometry), and caspase-3 activity.
Main Results:
- Survivin siRNA treatment led to near-complete suppression of survivin gene and protein expression in T24 cells.
- Significant reduction in cell growth, induction of G2/M phase arrest, and increased apoptosis were observed.
- Elevated caspase-3 activity, a marker of apoptosis, was detected following survivin inhibition.
Conclusions:
- RNA interference is an effective method for suppressing survivin expression in bladder cancer cells.
- Targeting survivin with siRNA demonstrates potential as a therapeutic approach to inhibit bladder cancer cell proliferation.
- Survivin-targeted siRNA therapy may serve as a valuable adjuvant treatment for bladder cancer.
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