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Updated: May 8, 2026
DNA Vector-based RNA Interference to Study Gene Function in Cancer
Published on: June 4, 2012
[Killing effect of double suicide genes mediated by lentivirus on lymphoma cells]
Yi-Rong Jiang1, Dao-Xin Ma, Xue-Liang Chen
1Department of Hematology, Qilu Hospital of Shandong University, Jinan, Shandong, 250012, PR China. jiangyirong1970@yahoo.com.cn
Background & Objective:
The lentiviral vectors can integrate interest genes into genome of the target cells that allow for stable transgenic expression even in non-dividing cells without evoking an immune response of the host. All the features have promised them to be used in vivo gene therapy. This study was designed to explore the killing effect of double suicide genes mediated by lentivirus on lymphoma cells (Raji).
Methods:
The three plasmids expressed lentivirus, packaging plasmid pCMV 8.2, envelope plasmid pCMV.VSVG and target plasmid (pHR'CS. GFP as control group, pHR'CS.CDglytk as experiment group) were packaged into 293T cells using lipofectine method. Supernatant was harvested and concentrated. The Raji cells were infected with the concentrated virus. The gene integration and expression were confirmed by fluorescence microscopy and RT-PCR. After prodrug GCV or/and 5-FC administration, MTT method was used to detect the growth inhibition rate (GIR) of Raji cells for evaluating the killing effect of CD and HSV-tk double suicide genes on Raji cells.
Results:
The three plasmids were effectively transferred into 293T cells. Green fluorescence on the cell was observed through fluorescence microscopy and a lot of virus particles were observed through transmission electronic microscopy. Double suicide genes mediated by lentivirus were effectively and stably expressed in Raji cells. The GIR of Raji cells using GCV or 5-FC was 51% or 50%, respectively, and it was apparently higher than that of untransfected cells(P< 0.01). When using GCV and 5-FC together, the GIR was 73%, which was apparently higher than that of group using GCV or 5-FC alone (P< 0.01).
Conclusion:
Double suicide genes mediated by lentiviral vector could transfect lymphoma cells effectively and stably. The double suicide gene system enhanced killing effect remarkably on lymphoma cells than CD/5FC or HSV-tk/GCV system alone.
Insights
Lentiviral vectors efficiently delivered double suicide genes into lymphoma cells, enhancing their killing effect. This gene therapy approach shows promise for treating lymphoma by combining GCV and 5-FC prodrugs for superior results.
Area of Science:
- Molecular Biology
- Gene Therapy
- Oncology
Context:
- Lentiviral vectors offer stable gene integration and expression in target cells, suitable for in vivo gene therapy.
- Lymphoma treatment remains a challenge, necessitating novel therapeutic strategies.
- Suicide gene therapy involves introducing genes that convert non-toxic prodrugs into toxic agents within cancer cells.
Purpose:
- To investigate the efficacy of a lentivirus-mediated double suicide gene system (cytosine deaminase and HSV-tk) against Raji lymphoma cells.
- To evaluate the gene integration and expression of the suicide genes in target cells.
- To assess the combined killing effect of the double suicide genes with GCV and 5-FC prodrugs.
Summary:
- Lentiviral vectors successfully transfected and stably expressed double suicide genes (cytosine deaminase and HSV-tk) in Raji lymphoma cells.
- Individual prodrugs (GCV or 5-FC) resulted in significant growth inhibition rates (51% and 50%, respectively) compared to controls.
- Combination therapy with both GCV and 5-FC achieved a 73% growth inhibition rate, significantly higher than monotherapy.
Impact:
- Demonstrates the potential of lentiviral vectors for delivering suicide genes in lymphoma therapy.
- Highlights the synergistic effect of combining cytosine deaminase/5-FC and HSV-tk/GCV systems for enhanced cancer cell killing.
- Provides a foundation for developing more effective gene-based treatments for B-cell lymphomas.
