Related Experiment Video
Updated: May 27, 2026
Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis
Published on: March 30, 2019
Biologically inhibitory effects of VEGF siRNA on endometrial carcinoma cells
Shuping Zhao1, Dehua Ma, Hongying Dai
1Department of Obstetrics and Gynecology, Affiliated Hospital of Qingdao University School of Medicine, Qingdao 266003, People's Republic of China. shuping.zhao@yahoo.com
Objective:
To study the biological effects of small inhibitory RNA (siRNA) on endometrial carcinoma cells through disrupting the mRNA of Vascular endothelial growth factor (VEGF), and its inhibitory functions on tumor cells' proliferation.
Methods:
The sequence-specific siRNA of VEGF165 and the mock siRNA were designed, they were transfected into the endometrial carcinoma cell line, Ishikawa cells. Total cell RNAs were extracted from the transfected cells. The mRNA levels of VEGF were then analyzed utilizing real-time fluorescence quantitative RT-PCR (FQ-PCR) post-transfection at 12, 24, 48, 72, and 168 h, respectively. Simultaneously its protein levels were also determined by western blot. Cell proliferations were monitored by MTT assay and cytopathological effects (CPE) were determined by Multispect Imaging System.
Result:
The mRNA and protein levels of VEGF declined post-transfection at 12, 24, 48, 72 h, respectively, with a maximum decrease post-transfection at 48 h. The proliferation of Ishikawa cells was significantly inhibited in a similar manner during this time window. All the data were repeated at least three independent times and similar results were achieved.
Conclusion:
The small siRNA of VEGF165 can effectively down-regulate its target mRNA and protein levels; consequently proliferation of Ishikawa cells was inhibited. The data strongly imply that VEGF may be involved in the genesis of endometrial carcinoma, which provides a new pathway to treat human endometrial caners.
Insights
Small interfering RNA (siRNA) targeting vascular endothelial growth factor (VEGF) effectively reduced VEGF mRNA and protein levels in endometrial carcinoma cells. This inhibition significantly suppressed tumor cell proliferation, suggesting VEGF
Area of Science:
- Molecular biology
- Oncology
- RNA interference
Background:
- Vascular Endothelial Growth Factor (VEGF) plays a crucial role in tumor angiogenesis and progression.
- Endometrial carcinoma is a significant gynecological malignancy.
- Targeting key growth factors offers a potential therapeutic strategy.
Purpose of the Study:
- To investigate the biological effects of small interfering RNA (siRNA) against VEGF in endometrial carcinoma cells.
- To assess the impact of VEGF mRNA disruption on tumor cell proliferation.
Main Methods:
- Design and transfection of sequence-specific VEGF siRNA into Ishikawa endometrial carcinoma cells.
- Quantification of VEGF mRNA and protein levels using real-time quantitative RT-PCR and Western blot.
- Assessment of cell proliferation and cytopathological effects via MTT assay and Multispect Imaging System.
Main Results:
- VEGF siRNA significantly reduced both VEGF mRNA and protein expression in Ishikawa cells, with maximum effect at 48 hours post-transfection.
- A corresponding significant inhibition of endometrial carcinoma cell proliferation was observed.
- Results were consistently reproducible across multiple independent experiments.
Conclusions:
- VEGF siRNA effectively downregulates VEGF expression at both mRNA and protein levels.
- Inhibition of VEGF leads to suppressed proliferation of endometrial carcinoma cells.
- VEGF is implicated in endometrial carcinoma development, presenting a potential therapeutic target.
Related Concept Videos
01:24Regulation of Angiogenesis and Blood Supply
02:34Inhibition of Cdk Activity
02:34Inhibition of CDK Activity
02:15Experimental RNAi
01:22MicroRNAs