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Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis
Published on: March 30, 2019
RNA interference-mediated gene silencing of vascular endothelial growth factor in colon cancer cells
Tie-Jun Li1, Jian-Ning Song, Kai Kang
1Department in General Surgery, the First Affiliated Hospital, Chongqing University of Medical Science, Chongqing 400016, China.
Aim:
To inhibit the expression of vascular endothelial growth factor (VEGF) in colon cancer cell line by RNA interference (RNAi).
Methods:
Followed the service of E-RNAi, we designed and constructed two kinds of shRNA expression vectors aiming at the VEGF gene, then transfected them into colon cancer HT29 cells by lipofectamine(TM) 2000. The level of VEGF mRNA was investigated by RT-PCR and Northern blotting. The protein expression of VEGF was observed by immunofluoresence staining and Western blotting.
Results:
We got two kinds of VEGF specific shRNA expression vectors which could efficiently inhibit the expression of VEGF in HT29 cells. RT-PCR, Northern blotting, immunofluoresence staining and Western blotting showed that inhibition rate for VEGF expression was up to 42%, 89%, 73% and 82%, respectively.
Conclusion:
The expression of VEGF can be inhibited by RNA interference in HT29 cells.
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