[Effects of vascular endothelial growth factor (VEGF) antisense oligodeoxynucleotide on mRNA and expression of VEGF, flt-1, and kinase insert domain containing receptor and VEGF excretion in human gallbladder carcinoma cells].
OBJECTIVE
To investigate the effects of vascular endothelial growth factor (VEGF) antisense oligodeoxynucleotide (ASODN) on the mRNA and protein expression of VEGF, Flt-1, and kinase insert domain containing receptor (KDR) and VEGF excretion in human gallbladder carcinoma cells.


METHODS
Human gallbladder carcinoma cells of the line GBC-SD were cultured and transfected with VEGF ASODN and sense oligodeoxynucleotide (SODN) mediated by Oligofectamine. The toxicity of SODN and Oligofectamine to the GBC-SD cells was examined by MTT method. RT-PCR was used to detect the mRNA and expression of VEGF, Flt-1, and KDR, and ELISA was used to detect the protein expression of VEGF.


RESULTS
MTT method showed that SODN and Oligofectamine were not toxic to the GBC-SD cells. The mRNA expression levels of VEGF, Flt-1, and KDR of the ASODN and ASODN + Oligofectamine groups were all significantly lower than those of the control group (all P < 0.05), and were the lowest 72 hours after transfection, and then gradually increased. ELISA showed that there were not significant differences in the VEGF protein concentration in the supernatant of the GBC-SD cells among the SODN, SODN + Oligofectamine, and control groups (all P < 0.05), however, the VEGF protein concentration in the supernatant of the GBC-SD cells of the ASDN and ASDN + Oligofectamine groups were significantly lower than that of the control group (both P < 0.05).


CONCLUSIONS
VEGF ASODN inhibits the mRNA and protein expression of VEGF, Flt-1, and KDR and VEGF excretion in human gallbladder carcinoma cells. Oligofectamine strengthens the effect of ASODN.
Fang
He-Qing
HQ
Department of Surgery, Second Affiliated Hospital, Medical School of Zhejiang University, Hangzhou 310009, China.
Li
Hai-Jun
HJ
Liu
Ying-Bin
YB
Wang
Xu-An
XA
Ma
Xiao-Ming
XM
Kong
Ying
Y
Chen
Yan
Y
Chen
De-Qin
DQ
Weng
Wei-Hong
WH
Zhang
Zhi-Ping
ZP
Devkota
K-R
KR
Wang
Jian-Wei
JW
Li
Jiang-Tao
JT
Cao
Li-Ping
LP
Peng
Shu-You
SY
D017209:Apoptosis; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005706:Gallbladder Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D006801:Humans; D020320:Oligoribonucleotides, Antisense / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis / Q000235:genetics*; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics
D020320:Oligoribonucleotides, Antisense; D012333:RNA, Messenger; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C501162:FLT1 protein, human; D040281:Vascular Endothelial Growth Factor Receptor-1; D040301:Vascular Endothelial Growth Factor Receptor-2