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Published on: March 30, 2019
Short interfering RNA-mediated gene silencing of vascular endothelial growth factor: effects on cellular
Abby L Mulkeen1, Teresa Silva, Peter S Yoo
1Department of Surgery, Yale Cancer Center, New Haven, Conn., USA.
Hypothesis:
By using short interfering RNA (siRNA) to inhibit the in vitro expression of vascular endothelial growth factor (VEGF) A, we hope to further investigate the presence of an autocrine loop in colon cancer cells. We hypothesize that VEGF inhibition will result in decreased cellular proliferation.
Design:
Human colon cancer cells were evaluated for the expression of VEGF and VEGF receptor 2 (VEGFR-2). In vitro assessments were then made of the ability of anti-VEGF siRNA to knock down expression of VEGF and the subsequent effect this decreased expression had on colon cancer cell proliferation.
Setting:
Surgical oncology research laboratory.
Interventions:
Human colon cancer cells from the RKO cell line were transfected with siRNA targeting the coding region of VEGF.
Main Outcome Measures:
Enzyme-linked immunosorbent assay, Northern blot analysis, and real-time quantitative polymerase chain reaction were performed to establish the ability of siRNA to decrease VEGF production. Proliferation assays were run on transfected and wild-type cells to establish concomitant decrease in VEGF expression and cellular proliferation.
Results:
The RKO colon cancer cells expressed both VEGF and VEGFR-2. Those cells transfected with siRNA targeting VEGF showed a 94% knockdown in VEGF expression and a 67% decrease in cellular proliferation.
Conclusion:
Colon cancer cells expressing VEGF and VEGFR-2 may possess an autocrine growth pathway that can be effectively targeted using RNA interference as an antiangiogenic therapy.
Insights
Short interfering RNA (siRNA) targeting vascular endothelial growth factor (VEGF) significantly reduced colon cancer cell proliferation. This suggests siRNA is a potential antiangiogenic therapy for colon cancers driven by VEGF autocrine loops.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Vascular Endothelial Growth Factor (VEGF) plays a crucial role in tumor angiogenesis and progression.
- Autocrine signaling loops involving VEGF may contribute to colon cancer cell growth.
- Targeting VEGF offers a potential therapeutic strategy for various cancers.
Purpose of the Study:
- To investigate the role of VEGF in colon cancer cell proliferation.
- To determine if inhibiting VEGF expression using short interfering RNA (siRNA) affects colon cancer cell growth.
- To explore the potential of siRNA as an antiangiogenic therapy for colon cancer.
Main Methods:
- Human colon cancer cells (RKO cell line) were analyzed for VEGF and VEGF Receptor 2 (VEGFR-2) expression.
- Cells were transfected with siRNA targeting VEGF.
- VEGF knockdown was confirmed using enzyme-linked immunosorbent assay, Northern blot, and quantitative PCR.
- Cell proliferation was assessed in transfected and wild-type cells.
Main Results:
- RKO colon cancer cells expressed both VEGF and VEGFR-2.
- siRNA transfection resulted in a 94% knockdown of VEGF expression.
- Colon cancer cell proliferation decreased by 67% following VEGF knockdown.
Conclusions:
- Colon cancer cells expressing VEGF and VEGFR-2 may utilize an autocrine growth pathway.
- RNA interference targeting VEGF can effectively inhibit colon cancer cell proliferation.
- VEGF inhibition via siRNA represents a promising antiangiogenic therapeutic approach for colon cancer.
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