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[Suppression of vascular endothelial growth factor expression by vector-based small interfering RNA in human tongue
Da-hai Yu1, Ying Cao, Zhi-wen Yao
1Dept. of Oral and Maxillofacial Surgery, College of Stomatology, Guangxi Medical University, Nanning 530021, China.
Objective:
To assess suppression effects of vector-based small interfering RNA (siRNA) on vascular endothelial growth factor (VEGF) expression of human tongue squamous carcinoma cell line (Tca8113) in vitro.
Methods:
Two siRNA targeting VEGF constructed in eukaryotic expression vector (Pu-VEGF-siRNA1, Pu-VEGF-siRNA2), eukaryotic expression vector as the experiment control, all of which were transfected into Tca8113 cells with Lipofectamine 2000. Non-transfection cell was used as negative control. VEGF mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry and enzyme linked immunosorbent assay (ELISA), respectively.
Results:
Compared to the experimental and negative controls, the expression of VEGF mRNA and protein were significantly decreased in the Pu-VEGF-siRNA1 group and Pu-VEGF-siRNA2 group. But there were no significant differences between two controls (P > 0.05).
Conclusion:
Vector-based siRNAs targeting VEGF are efficient in down-regulating VEGF expression in Tca8113 cells.
Insights
Vector-based small interfering RNA (siRNA) effectively reduced vascular endothelial growth factor (VEGF) expression in human tongue cancer cells. This study demonstrates siRNA
Area of Science:
- Molecular Biology
- Cancer Research
- Gene Silencing
Background:
- Vascular Endothelial Growth Factor (VEGF) plays a crucial role in tumor angiogenesis and progression.
- Overexpression of VEGF is implicated in the development and metastasis of various cancers, including tongue squamous cell carcinoma.
- Targeting VEGF offers a potential therapeutic strategy for cancer treatment.
Purpose of the Study:
- To evaluate the efficacy of vector-based small interfering RNA (siRNA) in suppressing VEGF expression.
- To investigate the in vitro effects of siRNA targeting VEGF on the human tongue squamous carcinoma cell line (Tca8113).
Main Methods:
- Construction of two siRNA molecules targeting VEGF within eukaryotic expression vectors (Pu-VEGF-siRNA1, Pu-VEGF-siRNA2).
- Transfection of Tca8113 cells using Lipofectamine 2000, with empty vector and non-transfected cells serving as controls.
- Quantification of VEGF mRNA and protein levels using reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry, and enzyme-linked immunosorbent assay (ELISA).
Main Results:
- Significant downregulation of both VEGF mRNA and protein expression was observed in cells transfected with Pu-VEGF-siRNA1 and Pu-VEGF-siRNA2 compared to controls.
- No significant difference in VEGF expression was found between the experimental control (empty vector) and the negative control (non-transfected cells).
Conclusions:
- Vector-based siRNAs targeting VEGF demonstrate significant efficiency in down-regulating VEGF expression in Tca8113 cells.
- These findings support the potential of vector-based siRNA technology as a therapeutic approach for tongue squamous cell carcinoma by inhibiting VEGF-driven pathways.
