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Updated: May 17, 2026
DNA Vector-based RNA Interference to Study Gene Function in Cancer
Published on: June 4, 2012
[Stable interference on P210(bcr/abl) gene expression by lentiviral vector-delivered shRNA in vitro and in vivo]
Yu-Feng Zhu1, Yuan-Zhan Wang, Fan-Yi Meng
1Department of Hematology, The Southern Medical University, Guangzhou, China.
Abstract:
P210(bcr/abl) fusion gene is indispensable for generation and progression of chronic myeloid leukemia (CML). Small molecule inhibitors, such as imatinib, are effective for P210(bcr/abl) gene mediated CML, but drug resistance may occur. The unique fusion junction of P210(bcr/abl) gene is an attractive target for therapeutic intervention using RNA interference (RNAi). This study was purposed to constructed the BaF3 cell line by viral vector which can stably express P210(bcr/abl) shRNA and P210(bcr/abl) mRNA at the same time, and investigate the effect of lentiviral-victor-delivered shRNA on P210(bcr/abl) gene expression. The infective rate of lentiviral vector on BaF3 cells with P210(bcr/abl) gene was assayed by fluorescent microscopy; the cell proliferation ability was determined by trypan blue exclusion; the P210(bcr/abl) mRNA and protein expressions were detected by RT-PCR and Western blot respectively. The results found that stable expression of the P210(bcr/abl) shRNA resulted in obvious inhibition of P210(bcr/abl) mRNA and protein expression and increased sensitivity of these P210(bcr/abl) gene transformed Ba/F3 cells to imatinib. The IC(50) to imatinib in these cells decreased < 50% as compared with Ba/F3-P210(bcr/abl) cells which did not express P210(bcr/abl) mRNA. The survival time of the lethal dose irradiated mice induced by intravenous injection of these Ba/F3 cells was longer than the other group induced by Ba/F3-P210(bcr/abl). It is concluded that stable expression of shRNA targeting the P210(bcr/abl) gene fusion junction may potentiate the effects of conventional therapy for CML.
Insights
This study developed a lentiviral vector to deliver short hairpin RNA (shRNA) targeting the P210 (bcr/abl) fusion gene, effectively inhibiting its expression in chronic myeloid leukemia (CML) cells. This approach enhances sensitivity to imatinib and improves survival in preclinical models.
Area of Science:
- Molecular Biology
- Gene Therapy
- Oncology
Context:
- The P210 (bcr/abl) fusion gene is a critical driver of chronic myeloid leukemia (CML).
- Imatinib therapy is effective but can be limited by drug resistance.
- RNA interference (RNAi) targeting the unique fusion junction offers a potential therapeutic strategy.
Purpose:
- To construct a BaF3 cell line stably expressing P210 (bcr/abl) shRNA using a lentiviral vector.
- To investigate the efficacy of lentiviral-delivered shRNA in inhibiting P210 (bcr/abl) gene expression.
- To evaluate the impact of P210 (bcr/abl) shRNA expression on cell proliferation, imatinib sensitivity, and in vivo survival.
Summary:
- Stable expression of P210 (bcr/abl) shRNA via lentiviral vector significantly reduced P210 (bcr/abl) mRNA and protein levels in BaF3 cells.
- Transduced cells exhibited increased sensitivity to imatinib, with a >50% decrease in IC50 values.
- Mice treated with shRNA-expressing cells showed prolonged survival compared to control groups.
Impact:
- Lentiviral-mediated shRNA delivery targeting the P210 (bcr/abl) fusion junction demonstrates potent gene silencing capabilities.
- This strategy enhances the efficacy of imatinib in preclinical models of CML.
- Stable shRNA expression offers a promising approach to potentiate conventional CML therapies and overcome drug resistance.
