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DNA Polymerases as targets for gene therapy of hepatocellular carcinoma
Hao Liu1, Qun Wei2, Jia Wang3,4
1Institute of Occupational Diseases, Zhejiang Academy of Medical Sciences, 182 Tianmushan Road, Hangzhou, 310013, Zhejiang, P.R. China. liuh_1985@163.com.
Background:
Hepatocyte carcinoma (HCC) is one of the most common malignancies worldwide. Despite many achievements in diagnosis and treatment, HCC mortality remains high due to the malignant nature of the disease. Novel approaches, especially for targeted therapy, are being extensively explored. Gene therapy is ideal for such purpose for its specific expression of exogenous genes in HCC cells driven by tissue-specific promoter. However strategies based on correction of mutations or altered expressions of genes responsible for the development/progression of HCC have limitations because these aberrant molecules are not presented in all cancerous cells. In the current work, we adopted a novel strategy by targeting the DNA replication step which is essential for proliferation of every cancer cell.
Methods:
A recombinant adenovirus with alpha fetoprotein (AFP) promoter-controlled expressions of artificial microRNAs targeting DNA polymerases α, δ, ε and recombinant active Caspase 3, namely Ad/AFP-Casp-AFP-amiR, was constructed.
Results:
The artificial microRNAs could efficiently inhibit the expression of the target polymerases in AFP-positive HCC cells at both RNA and protein levels, and HCC cells treated with the recombinant virus Ad/AFP-Casp-AFP-amiR exhibited significant G0/1 phase arrest. The proliferation of HCC cells were significantly inhibited by Ad/AFP-Casp-AFP-amiR with increased apoptosis. On the contrary, the recombinant adenovirus Ad/AFP-Casp-AFP-amiR did not inhibit the expression of DNA polymerases α, δ or ε in AFP-negative human normal liver cell HL7702, and showed no effect on the cell cycle progression, proliferation or apoptosis.
Conclusions:
Inhibition of DNA polymerases α, δ and ε by AFP promoter-driven artificial microRNAs may lead to effective growth arrest of AFP-positive HCC cells, which may represent a novel strategy for gene therapy by targeting the genes that are essential for the growth/proliferation of cancer cells, avoiding the limitations set by any of the individually altered gene.
Insights
Hepatocellular carcinoma (HCC) gene therapy effectively targets DNA replication in cancer cells using artificial microRNAs. This novel approach inhibits proliferation and induces apoptosis in AFP-positive HCC cells, offering a promising new treatment strategy.
Area of Science:
- Oncology
- Gene Therapy
- Molecular Biology
Background:
- Hepatocellular carcinoma (HCC) remains a significant global health challenge with high mortality rates despite advances in treatment.
- Current targeted therapies for HCC face limitations due to the heterogeneity of genetic alterations in cancer cells.
- A novel strategy targeting essential cellular processes, such as DNA replication, is needed for effective HCC treatment.
Purpose of the Study:
- To develop and evaluate a novel gene therapy strategy for hepatocellular carcinoma (HCC).
- To target DNA replication, a fundamental process for cancer cell proliferation, in HCC.
- To utilize a tissue-specific promoter for targeted delivery of therapeutic agents to HCC cells.
Main Methods:
- Construction of a recombinant adenovirus (Ad/AFP-Casp-AFP-amiR) expressing artificial microRNAs against DNA polymerases and active Caspase 3, driven by the alpha fetoprotein (AFP) promoter.
- Assessment of the inhibition of DNA polymerases α, δ, and ε expression in HCC cells at both RNA and protein levels.
- Evaluation of the effects of the recombinant adenovirus on HCC cell cycle progression, proliferation, and apoptosis, as well as on normal liver cells.
Main Results:
- The artificial microRNAs efficiently inhibited target DNA polymerase expression in AFP-positive HCC cells.
- Ad/AFP-Casp-AFP-amiR treatment resulted in significant G0/1 phase arrest, inhibited proliferation, and increased apoptosis in HCC cells.
- The recombinant adenovirus showed no significant effects on AFP-negative normal liver cells, indicating specificity.
Conclusions:
- Targeting DNA polymerases α, δ, and ε via AFP promoter-driven artificial microRNAs offers an effective strategy for growth arrest in AFP-positive HCC cells.
- This approach represents a novel gene therapy for HCC by targeting universally essential genes for cancer proliferation.
- The strategy overcomes limitations of therapies targeting individually altered genes in HCC.
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