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Updated: Aug 8, 2026
An Orthotopic Bladder Cancer Model for Gene Delivery Studies
Published on: December 2, 2013
[Multiple inhibitory mechanisms of wild-type p16 gene transfer into human bladder cells by retroviral vector]
Objective:
To investigate the inhibitory mechanisms of wild-type p16 gene transfer into human bladder cancer cells with or without endogenous p16 gene expression.
Methods:
p16 recombinant retrovirus vector was constructed and transfected in human bladder cancer cell EJ and 253J. Northern blot analysis and immunocytochemistry were used to detect the expression of exogenous p16 gene. Flow cytometry analysis was used to detect tumor cell cycle. Electronic microscope and in situ labelling apoptotic DNA fragment detection (TUNEL) was used to detect tumor cell apoptosis after gene transfection. Tumorigenicity was observed in nude mice.
Results:
The growth rate of the transfected EJ cells was significantly retarded, the majority of transfected EJ cells were arrested on G(0) + G(1) phases of cell cycle, tumorigenicity of the transfected cells was reduced on nude mice. H-ras gene expression in transfected EJ cells reduced as compared with non-transfection. Apoptotic cells were observed in transgenic 253J cells.
Conclusions:
Malignant proliferation of bladder cancer cells can be inhibited by exogenous p16 gene transfection via different molecular pathway.
Insights
Wild-type p16 gene transfer inhibits bladder cancer cell growth and tumorigenicity. This exogenous p16 gene transfection impacts cell cycle and induces apoptosis through distinct molecular pathways.
Area of Science:
- Molecular biology
- Cancer genetics
- Gene therapy
Context:
- Bladder cancer is a significant health concern.
- Understanding p16 gene's role in tumor suppression is crucial.
- Investigating gene therapy for bladder cancer treatment.
Purpose:
- To explore the inhibitory effects of wild-type p16 gene transfer.
- To elucidate the mechanisms of p16 gene therapy in bladder cancer cells.
- To assess p16 gene expression in cells with and without endogenous p16.
Summary:
- Wild-type p16 gene was transfected into human bladder cancer cell lines (EJ and 253J) using a retrovirus vector.
- Exogenous p16 gene expression was confirmed via Northern blot and immunocytochemistry.
- Transfected cells showed reduced proliferation, G(0)-G(1) cell cycle arrest, decreased tumorigenicity in mice, reduced H-ras expression, and induced apoptosis.
Impact:
- Demonstrates the potential of p16 gene therapy for bladder cancer.
- Highlights differing molecular pathways influenced by p16 gene transfer.
- Provides insights into targeting cell cycle and apoptosis for cancer treatment.
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