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Published on: January 22, 2021
[Effect of siRNA targeting c-Myc and VEGF on human colorectal cancer cells]
Jian-dong Tai1, Guang-yi Wang, Tong-jun Liu
1Department of Colorectal Surgery, The First Hospital, Jilin University, Changchun, China.
Objective:
To investigate the biological behavioral effects of specific siRNA expression plasmids targeted against c-Myc and vascular endothelial growth factor (VEGF) on human colorectal cancer cell line Volo.
Methods:
The expression plasmids with small interfering RNA (siRNA) aiming at c-Myc and VEGF were designed and constructed respectively, then transfected into Volo cells by eukaryocyte transfection technique. The protein expressions of c-Myc and VEGF were detected by Western blotting. Cellular proliferation, apoptosis, cycle distribution and invasion character were analyzed by tetrazolium bromide colorimetry (MTT), flow cytometry (FCM), TUNEL assay and matrigel invasion assay respectively.
Results:
Enzymatic digestion and DNA sequencing confirmed that the c-Myc and VEGF specific siRNA expression plasmids were constructed successfully. After plasmids were transfected into cells, the protein expressions of c-Myc and VEGF were significantly down-regulated respectively as compared with control group (P<0.01). The cellular proliferation inhibitory rates in c-Myc siRNA group, VEGF siRNA group and c-Myc+VEGF group were (59.20+/-5.05)%, (32.31+/-3.48)% and (75.81+/-7.89)% respectively, which were higher than that in control group [(6.80+/-1.45)%] (all P<0.05). The cell apoptosis rate in above 3 groups were (40.50+/-4.37)%, (21.30+/-2.98)% and (62.59+/-9.66)% respectively, which were higher than that in control group [(2.90+/-0.36)%] (all P<0.05). The cell invasion rates in VEGF siRNA group and c-Myc+VEGF siRNA group were (7.34+/-3.65)% and (2.80+/-1.02)%, which were lower than that in control group [(18.57+/-7.46)%] (P<0.05). The effect of c-Myc+VEGF siRNA group was greater.
Conclusions:
The specific siRNA efficiently silences the expression of c-Myc and VEGF, subsequently, suppresses the cell proliferation, triggers the cell apoptosis and inhibits the cell invasiveness in these transfected colorectal cancer Volo cells. In addition, the synergism of siRNA-c-Myc and siRNA-VEGF in transfected cells can be found.
Insights
Specific siRNA targeting c-Myc and vascular endothelial growth factor (VEGF) effectively suppressed colorectal cancer cell proliferation and invasion. Combined siRNA treatment demonstrated synergistic effects, enhancing apoptosis and inhibiting tumor growth in Volo cells.
Area of Science:
- Molecular Biology
- Oncology
- Gene Silencing
Context:
- Colorectal cancer (CRC) remains a significant global health challenge.
- Targeting key oncogenes like c-Myc and growth factors such as vascular endothelial growth factor (VEGF) is a promising therapeutic strategy.
- Small interfering RNA (siRNA) offers a precise method for gene expression modulation.
Purpose:
- To evaluate the efficacy of specific siRNA expression plasmids targeting c-Myc and VEGF.
- To investigate the biological and behavioral effects of these siRNAs on the human colorectal cancer cell line Volo.
- To assess the synergistic effects of combined c-Myc and VEGF siRNA delivery.
Summary:
- Successful construction and transfection of c-Myc and VEGF siRNA expression plasmids into Volo cells were confirmed.
- siRNA-mediated silencing significantly downregulated c-Myc and VEGF protein expression.
- Treatment with c-Myc siRNA, VEGF siRNA, or both notably inhibited cell proliferation, increased apoptosis, and reduced cell invasion, with combined siRNA showing enhanced effects.
Impact:
- Demonstrates the potent anti-cancer activity of siRNA targeting c-Myc and VEGF in colorectal cancer models.
- Highlights the potential synergistic therapeutic benefit of simultaneously silencing c-Myc and VEGF.
- Provides a foundation for developing novel RNA-based therapies for colorectal cancer.
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