RNA interference-mediated gene silencing of vascular endothelial growth factor in colon cancer cells.
OBJECTIVE
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.


CONCLUSIONS
The expression of VEGF can be inhibited by RNA interference in HT29 cells.
Li
Tie-Jun
TJ
Department in General Surgery, the First Affiliated Hospital, Chongqing University of Medical Science, Chongqing 400016, China.
Song
Jian-Ning
JN
Kang
Kai
K
Tong
Shu-Sheng
SS
Hu
Zan-Lan
ZL
He
Tong-Chuan
TC
Zhang
Bing-Qiang
BQ
Zhang
Cai-Quan
CQ
D000230:Adenocarcinoma / Q000235:genetics* / Q000378:metabolism; D015152:Blotting, Northern; D015153:Blotting, Western; D003110:Colonic Neoplasms / Q000235:genetics* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D020868:Gene Silencing / Q000187:drug effects*; D005822:Genetic Vectors / Q000235:genetics; D019073:HT29 Cells; D006801:Humans; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism
D012333:RNA, Messenger; D034741:RNA, Small Interfering; D042461:Vascular Endothelial Growth Factor A