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[The experimental study of transfected sFlt-Ig gene on K562 leukemia cell growth in nude mice]
1Institute of Hematology, CAMS & PUMC, State key Lab. of Experimental Hematology, Tianjin 300020, China.
Objective:
To observe the influence of transfected soluble vascular endothelial growth factor (VEGF) receptor (sFlt-1) gene on K562 leukemia cell growth in vivo.
Methods:
(1) The binding region of VEGF receptor (Flt-1) ligand was combined with fragment of IgH stable region to construct Flt-Ig fusion gene and insert into pcDNA3 vector. (2) By using electroporation, the pcDNA3/Flt-Ig was transfected into K562 leukemia cells, and selected by G418. Flt-Ig mRNA expression was detected by RT-PCR. (3) The transfected pcDNA3/Flt-Ig and pcDNA3-Ig K562 cells were respectively transplanted into nude mice and the tumor volume was dynamically measured.
Results:
Five subclones of K562 cells with high expression of Flt-Ig gene have been established, one of them was transplanted into 6 nude mice. The tumor volume of experimental mice was obviously smaller than that of control mice, about one half of the control group (P < 0.05).
Conclusion:
The growth of transfected pcDNA3/Flt-Ig K562 cells was significantly inhibited. It is possible that soluble Flt-Ig protein secreted from K562/Flt-Ig cells neutralized VEGF produced from tumor cells, therefore inhibited the tumor angiogenesis.
Insights
Transfecting K562 leukemia cells with the soluble vascular endothelial growth factor (VEGF) receptor (sFlt-1) gene significantly inhibited tumor growth in mice. This suggests sFlt-1 may block tumor angiogenesis by neutralizing VEGF.
Area of Science:
- Molecular biology
- Cancer research
- Gene therapy
Context:
- K562 leukemia cells are a model for chronic myeloid leukemia.
- Vascular Endothelial Growth Factor (VEGF) plays a crucial role in tumor angiogenesis.
- Soluble VEGF receptors can act as antagonists to inhibit VEGF signaling.
Purpose:
- To investigate the effect of transfecting K562 leukemia cells with the soluble VEGF receptor (sFlt-1) gene on tumor growth in vivo.
- To construct and express a fusion gene encoding sFlt-1 in K562 cells.
- To assess the impact of sFlt-1 expression on tumor development in a mouse model.
Summary:
- A fusion gene (Flt-Ig) encoding the soluble VEGF receptor was constructed and transfected into K562 leukemia cells.
- Transfected cells (K562/Flt-Ig) showed stable expression of the Flt-Ig gene.
- Tumor growth in nude mice implanted with K562/Flt-Ig cells was significantly reduced compared to control groups.
Impact:
- The study demonstrates that introducing the sFlt-1 gene into leukemia cells can inhibit their growth.
- This suggests a potential therapeutic strategy for leukemia by targeting tumor angiogenesis.
- Soluble Flt-Ig protein secreted by transfected cells may neutralize tumor-derived VEGF, thereby inhibiting angiogenesis.