[The inhibitory effect of vascular endothelial growth factor antisense oligonucleotides in angiogenesis of thyroid carcinoma].
OBJECTIVE
To investigate the inhibitive effect of antisense oligonucleotide (ASODN) on vascular endothelial growth factor (VEGF) expression and endothelial cell growth in thyroid carcinoma.


METHODS
Targeted ASODN of VEGF was designed and synthesized, then transfected to TT (medullary thyroid carcinoma) cell line and the culture supernatant was collected in which ECV304 (endothelial cell line) was seeded. At the same time positive control [sense oligonucleotides (SODN) group] and normal control were set for comparison. Cell growth condition was observed under microscope. RT-PCR and immuocytochemistry were used for detection of VEGF mRNA and protein expression in TT cells. MTT assay was used for cell growth inhibition ratio (IR) of TT and ECV304 cells, flow cytometry (FCM) for apoptotic index (AI) of ECV304 cells and acridine orange/ethidium bromide (AO/EB) staining for apoptotic morphology of ECV304 cells.


RESULTS
As compared with positive and normal control groups, VEGF mRNA and protein expressions in TT cells of ASODN transfection groups were obviously decreased (P < 0.01). Cell growth was not influenced apparently in ECV304 cells with direct ASODN administration, but ECV304 cell growth in ASODN conditioned medium was significantly inhibited and IR (0.21 +/- 0.03, 0.31 +/- 0.01, 0.42 +/- 0.22) was significantly higher than that of SODN group (0.05 +/- 0.03, P < 0.01), with the presence of apparent apoptosis. The effect mentioned above was in a dose-dependent manner.


CONCLUSIONS
ASODN can suppress endothelial cell growth and inhibit tumor angiogenesis possibly by specifically blocking VEGF expression in thyroid carcinoma.
Zhang
Hai-yan
HY
Department of Endocrinology, the First Affiliated Hospital, China Medical University, Shenyang 110001, China.
Gao
Da-xin
DX
Li
Ping
P
Ren
Lu-ping
LP
Cao
Cui-ping
CP
Liu
Guo-liang
GL
D017209:Apoptosis; D018276:Carcinoma, Medullary / Q000098:blood supply* / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D004730:Endothelium, Vascular / Q000166:cytology; D006801:Humans; D009389:Neovascularization, Pathologic / Q000517:prevention & control*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics; D013964:Thyroid Neoplasms / Q000098:blood supply* / Q000378:metabolism* / Q000473:pathology; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*
D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A