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Updated: Sep 20, 2026
An Orthotopic Bladder Cancer Model for Gene Delivery Studies
Published on: December 1, 2013
Growth inhibiting effects of antisense eukaryotic expression vector of proliferating cell nuclear antigen gene on
Qiangsong Tong1, Fuqing Zeng, Chen Lin
1Department of Urology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
Objective:
To explore the growth inhibiting effects on human bladder cancer by antisense RNA targeting the proliferating cell nuclear antigen (PCNA) gene.
Methods:
The eukaryotic expression vector for antisense PCNA cDNA was constructed and transferred into a bladder cancer EJ cell line. The PCNA expression in the cancer cells was detected by RT-PCR and Western blotting assays. The in vitro proliferation activities of the transferred cells were observed by growth curve, tetrazolium bromide (MTT) colorimetry, tritiated thymidine ((3)H-TdR) incorporation, flow cytometry and clone formation testing, while its in vivo anti-tumor effects were detected on nude mice allograft models.
Results:
After the antisense vector, pLAPSN, was transferred, cellular PCNA expression was inhibited at both protein and mRNA levels. The growth rates of EJ cells were reduced from 27.91% to 62.07% (P < 0.01), with an inhibition of DNA synthesis rate by 52.31% (P < 0.01). Transferred cells were blocked at G(0)/G(1) phases in cell-cycle assay, with the clone formation ability decreased by 50.81% (P < 0.01). The in vivo carcinogenic abilities of the transferred cancer cells were decreased by 54.23% (P < 0.05).
Conclusions:
Antisense PCNA gene transfer could inhibit the growth of bladder cancer cells in vitro and in vivo, which provided an ideal strategy for gene therapy of human cancers.
Insights
Antisense RNA targeting the proliferating cell nuclear antigen (PCNA) gene inhibited human bladder cancer growth. This gene therapy strategy showed significant in vitro and in vivo anti-tumor effects.
Area of Science:
- Oncology
- Molecular Biology
- Gene Therapy
Background:
- Human bladder cancer remains a significant health concern.
- Targeting key cellular proliferation markers is a promising therapeutic avenue.
- Proliferating cell nuclear antigen (PCNA) is crucial for DNA replication and cell cycle progression.
Purpose of the Study:
- To investigate the efficacy of antisense RNA targeting the proliferating cell nuclear antigen (PCNA) gene for inhibiting human bladder cancer growth.
- To evaluate the in vitro and in vivo anti-tumor effects of PCNA gene silencing in bladder cancer cells.
Main Methods:
- Construction and transfer of a eukaryotic expression vector encoding antisense PCNA cDNA into human bladder cancer EJ cells.
- Assessment of PCNA expression using RT-PCR and Western blotting.
- Evaluation of in vitro proliferation using growth curves, MTT assays, (3)H-TdR incorporation, flow cytometry, and clone formation assays.
- Assessment of in vivo anti-tumor effects in nude mice allograft models.
Main Results:
- Successful inhibition of PCNA expression at both mRNA and protein levels in transferred EJ cells.
- Significant reduction in EJ cell growth rates (27.91%–62.07%) and DNA synthesis (52.31%).
- Cell cycle arrest at the G(0)/G(1) phase, decreased clone formation ability (50.81%), and reduced in vivo tumor formation (54.23%).
Conclusions:
- Antisense PCNA gene transfer effectively inhibits human bladder cancer cell proliferation both in vitro and in vivo.
- This approach represents a potential gene therapy strategy for human cancers.
- Targeting PCNA offers a viable pathway for developing novel anti-cancer treatments.
