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.
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.