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Updated: Jun 29, 2025
3-D Cell Culture System for Studying Invasion and Evaluating Therapeutics in Bladder Cancer
Published on: September 13, 2018
Hsa_circ_0005320 affects cell proliferation and the cell cycle via the IGF2BP3/CDK2 axis in bladder cancer
Quansong Ji1, Feilu Ma2, Xiling Zhang1
1Department of Urology, the Fourth Affiliated Hospital of China Medical University, Shenyang, Liaoning, China.
Background:
Circular RNAs (circRNAs), which are covalently closed non-coding RNAs, are frequently dysregulated in cancer. However, their precise role in bladder cancer (BCa) remains largely unknown.
Methods:
Expression of hsa_circ_0005320 in tissues and cell lines was detected using quantitative real-time PCR. Proliferation and colony forming capacity of BCa cells were assessed using Cell Counting Kit-8, ethynyl-labeled deoxyuridine, and colony formation assays. The cell cycle was analyzed using flow cytometry. Protein expression of insulin-like growth factor II mRNA-binding protein 3 (IGF2BP3) and cyclin dependent kinase 2 (CDK2) was examined using western blots. The binding of RNA and protein was validated using RNA immunoprecipitation. Additionally, xenograft tumor models were established to validate the function of hsa_circ_0005320 in vivo.
Results:
We screened hsa_circ_0005320 from previous high-throughput sequencing and found that it was highly expressed in BCa tissues and associated with tumor differentiation and depth of invasion in BCa patients. Through functional experiments, we demonstrated that hsa_circ_0005320 promoted cell proliferation and regulated the cell cycle. Mechanistically, hsa_circ_0005320 interacted with and upregulated the expression of IGF2BP3, which binds to and enhances the stability of CDK2 mRNA. Furthermore, knockdown of hsa_circ_0005320 resulted in a reduction in tumor burden in vivo.
Conclusions:
Collectively, these findings highlight the pro-oncogenic role of hsa_circ_0005320 in BCa through the IGF2BP3/CDK2 axis, providing valuable insights into the mechanism of circRNAs in tumor progression.
Insights
Circular RNAs (circRNAs) promote bladder cancer (BCa) by increasing cell proliferation via the IGF2BP3/CDK2 pathway. Targeting hsa_circ_0005320 may offer a new therapeutic strategy for BCa.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Circular RNAs (circRNAs) are non-coding RNAs with dysregulated expression in various cancers.
- The specific role of circRNAs in bladder cancer (BCa) pathogenesis is not well understood.
Purpose of the Study:
- To investigate the role and mechanism of hsa_circ_0005320 in bladder cancer.
- To elucidate the molecular pathway involving hsa_circ_0005320 in BCa progression.
Main Methods:
- Quantitative real-time PCR to detect hsa_circ_0005320 expression.
- Cell proliferation, colony formation, and cell cycle assays to assess BCa cell behavior.
- Western blotting and RNA immunoprecipitation to analyze protein-RNA interactions.
- Xenograft models to evaluate in vivo tumor growth.
Main Results:
- hsa_circ_0005320 was highly expressed in BCa tissues and correlated with poor differentiation and deep invasion.
- hsa_circ_0005320 promoted BCa cell proliferation and cell cycle progression.
- hsa_circ_0005320 upregulated insulin-like growth factor II mRNA-binding protein 3 (IGF2BP3), enhancing cyclin dependent kinase 2 (CDK2) mRNA stability.
- Knockdown of hsa_circ_0005320 reduced tumor burden in vivo.
Conclusions:
- hsa_circ_0005320 acts as a pro-oncogenic factor in bladder cancer.
- The IGF2BP3/CDK2 axis is a key mechanism through which hsa_circ_0005320 drives tumor progression.
- hsa_circ_0005320 represents a potential therapeutic target for bladder cancer.
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