[Influence of nuclear factor-kappaB decoy transfection on nuclear factor-kappaB activity and vascular endothelial growth factor/urokinase-type plasminogen activator/intercellular cell adhesion molecule-1 level of SKOV3 cells].
OBJECTIVE
To investigate the effect of nuclear factor-kappaB (NF-kappaB) decoy oligodeoxynucleotides (ODN) on NF-kappaB activation and expressions of vascular endothelial growth factor (VEGF), urokinase-type plasminogen activator (uPA), intercellular cell adhesion molecule-1 (ICAM-1) of SKOV3 cell.


METHODS
After transfected with NF-kappaB decoy ODN, SKOV3 cells were stimulated by IL-1beta and cultured for 6, 12, 24, and 48 h, respectively. NF-kappaB DNA binding activity was measured by gel mobility shift assay and mRNA levels of ICAM-1, uPA, VEGF were analyzed by RT-PCR and VEGF level in supernatant was detected by enzyme-linked immunosorbent assay (ELISA).


RESULTS
(1) SKOV3 cells expressed constitutive NF-kappaB DNA binding activity and stimulation of IL-1beta resulted in a significant increase of the binding activity. NF-kappaB decoy ODN transfection significantly decreased constitutive level and elevated level of NF-kappaB DNA binding activity by IL-1beta at 6, 12, 24, and 48 h in vitro (P < 0.05). (2) The constitutive and activated mRNA levels of uPA, VEGF, ICAM-1 and VEGF levels in the supernatant were significantly inhibited by NF-kappaB decoy ODN transfection especially at 6 h after transfection in vitro.


CONCLUSIONS
Both NF-kappaB DNA binding activity and expressions of uPA, VEGF, and ICAM-1 at 48 h were upregulated by IL-1beta, and inhibited by transfection with NF-kappaB decoy ODN in SKOV3 cell line. NF-kappaB decoy ODN transfection shows promise as a novel molecular approach for ovarian cancer.
Lu
Jian-ping
JP
Department of Obstetrics and Gynecology Hospital, Fudan University, Shanghai 200011, China.
Sun
Hong
H
Cao
Chang-chun
CC
Ou
Zhou-luo
ZL
D018284:Cystadenocarcinoma, Serous / Q000378:metabolism / Q000473:pathology; D005260:Female; D015316:Genetic Therapy; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis / Q000235:genetics; D016328:NF-kappa B / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology*; D010051:Ovarian Neoplasms / Q000378:metabolism* / Q000473:pathology; D014162:Transfection; D014407:Tumor Cells, Cultured; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis / Q000235:genetics; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis* / Q000235:genetics
D016328:NF-kappa B; D009838:Oligodeoxyribonucleotides; D042461:Vascular Endothelial Growth Factor A; D018799:Intercellular Adhesion Molecule-1; D014568:Urokinase-Type Plasminogen Activator