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The chimeric multi-domain proteins mediating specific DNA transfer for hepatocellular carcinoma treatment
Encheng Yang1, Xiao Li2, Ningyi Jin2
1Department of Gastroenterology and Hepatology, The Second Affiliated Hospital of Harbin Medical University, Harbin, 150086 China.
Aim:
This study was aimed to evaluate the therapeutic efficiency of a non-virus based specific chimeric multi-domain DNA transferred with apoptin in human hepatocellular carcinoma (HCC) HepG-2 cells in vitro and in mice H22 cells in vivo.
Methods:
We firstly constructed the multi-domain recombinant chimeric proteins based on recombinant proteins [G (yeast GAL4), NG (none GAL4), TG (GAL4 + Tat protein) and TNG (Tat protein)] and pUAS-Apoptin plasmid, and transfected them into human HepG-2 cells. The antitumor effect of this multi-domain recombinant chimeric proteins to HCC cells were detected by MTT assay, AO/EB staining, DAPI staining and Annexin V assay. In order to find the pathway of cell apoptosis, the Caspase (1, 3, 6 and 8) activity was detected. We then constructed the H22 liver cancer mice model and analyzed the anti-tumor rate and mice survival rate after treated with G/pUAS-Apoptin NG/pUAS-Apoptin TG/pUAS-Apoptin, and TNG/pUAS-Apoptin.
Results:
MTT results showed that the Tat protein (TG and TNG) significantly induced cell death in a time dependent manner. AO/EB, DAPI, Annexin V and Caspases assay results indicated that the Caspase 1, 3, 6 and 8 were highly expressed in TG/pUAS-Apoptin, and TNG/pUAS-Apoptin treated mouse groups. The antitumor rate and survival rate in TG/pUAS-Apoptin, and TNG/pUAS-Apoptin treated mouse groups were higher than in the other groups.
Conclusion:
The Tat-apoptin is a potential anti-tumor agent for HCC treatment with remarkable anti-tumor efficacy and high safety based on non-virus gene transfer system. The anti-tumor function may be associated with high expression of Caspase 1, 3, 6 and 8.
Insights
Tat-apoptin, a non-virus gene transfer system, shows significant anti-tumor efficacy against hepatocellular carcinoma (HCC) in vitro and in vivo. This novel approach demonstrates high safety and potential for HCC treatment by inducing apoptosis via Caspase activation.
Area of Science:
- Oncology
- Gene Therapy
- Molecular Biology
Background:
- Hepatocellular carcinoma (HCC) is a major global health concern.
- Current HCC treatments have limitations in efficacy and safety.
- Novel therapeutic strategies are needed for effective HCC management.
Purpose of the Study:
- To evaluate the therapeutic efficiency of a non-virus based chimeric multi-domain DNA transferred with apoptin in HCC.
- To assess the in vitro efficacy in human HCC HepG-2 cells and in vivo efficacy in mice H22 cells.
- To investigate the underlying mechanism of action, including apoptosis pathways.
Main Methods:
- Construction of multi-domain recombinant chimeric proteins with apoptin.
- Transfection into human HepG-2 cells and H22 liver cancer mice models.
- Assessment of antitumor effects using MTT assays, AO/EB, DAPI, Annexin V staining, and Caspase activity detection.
Main Results:
- Tat protein (TG and TNG) significantly induced HCC cell death in a time-dependent manner.
- TG/pUAS-Apoptin and TNG/pUAS-Apoptin treatments led to high expression of Caspase 1, 3, 6, and 8.
- Enhanced antitumor rates and survival rates were observed in mice treated with TG/pUAS-Apoptin and TNG/pUAS-Apoptin.
Conclusions:
- Tat-apoptin, delivered via a non-virus system, exhibits significant anti-tumor efficacy and safety for HCC treatment.
- The anti-tumor function is potentially mediated by the upregulation of Caspase 1, 3, 6, and 8.
- This non-virus gene transfer system represents a promising therapeutic agent for HCC.
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