Colorectal Cancer
Copyright ©2007 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Oct 28, 2007; 13(40): 5312-5316
Published online Oct 28, 2007. doi: 10.3748/wjg.v13.i40.5312
RNA interference-mediated gene silencing of vascular endothelial growth factor in colon cancer cells
Tie-Jun Li, Jian-Ning Song, Kai Kang, Shu-Sheng Tong, Zan-Lan Hu, Tong-Chuan He, Bing-Qiang Zhang, Cai-Quan Zhang
Tie-Jun Li, Kai Kang, Bing-Qiang Zhang, Cai-Quan Zhang, The First Affiliated Hospital, Chongqing University of Medical Sciences, Chongqing 400016, China
Jian-Ning Song, Zan-Lan Hu, Shu-sheng Tong, The Ninth Hospital of Chongqing, Chongqing 400700, China
Tong-Chuan He, Molecular Oncology Laboratory, University of Chicago Medical Center, IL 60637, United States
Author contributions: All authors contributed equally to the work.
Correspondence to: Professor Cai-Quan Zhang, Department in General Surgery, the First Affiliated Hospital, Chongqing University of Medical Science, Chongqing 400016,China. cqltj001@163.com
Telephone: +86-13983764504
Received: March 28, 2007
Revised: August 10, 2007
Accepted: August 24, 2007
Published online: October 28, 2007
Abstract

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 lipofectamineTM 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.

Keywords: RNA interference, Vascular endothelial growth factor, Colon cancer, Northern blotting, Western blotting