Background & objective: It had been reported that nuclear factor kappa b (NF kappa B) was associated with in vitro angiogenesis, but its role in angiogenesis of human ovarian carcinoma (HOC) had not been reported yet. The objective of this paper was to investigate the role and the mechanism of NF kappa B in HOC angiogenesis in chick chorioallantcic membrane (CAM).
Methods: The HOC TYK cells were implanted into 10-day CAM by 1 x 10(9)/per egg to establish the HOC CAM model. (1) With 20 eggs, the levels of NF kappa B, integrin alpha V beta 3, basic fibroblast growth factor (bFGF) at 0 h, 6 h, 12 h, 24 h, 48 h, 72 h time point after cells implanted were determined with ELISA. (2) The relationship among NF kappa B, bFGF,alpha V beta 3 and the effect of them on HOC angiogenesis in CAM were determined. Another 80 eggs were divided into 6 groups, normal saline (NS) (control group, 16 eggs), anti- NF kappa B (A group, 16 eggs), anti-bFGF (B group, 16 eggs), anti-NF kappa B + anti-bFGF(D group, 8 eggs) were added respectively into CAM at 6h time point after the cells implanted, and then anti-integrin alpha V beta 3 (C group, 16 eggs), anti-NF kappa B +anti-alpha V beta 3 (E group, 8 eggs) antibodies were added at 12h time point. Eight eggs from NS, A, B, C groups were taken out to determine the levels of NF kappa B, alpha V beta 3, bFGF at 48 h after the antibodies were added. The other eggs were incubated up to 5 days, and the vessel area/area were determined with image analysis.
Results: (1)The levels of NF kappa B and bFGF increased significantly at 6h, and that of V beta 3 increase significantly at 12 h time point after implantation (P< 0.01) (0.185+/-0.01, 0.771+/-0.16, 0.231+/-0.02), the peaks of the three levels were reached at 48 h (P< 0.01) (0.337+/-0.01, 1.639+/-0.01, 0.349+/-0.01). The levels of bFGF was significantly higher than those of NF kappa B and alpha V beta 3 at each time point (P< 0.01), but there were no significant difference between NF kappa B and alpha V beta 3 (P >0.05). (2)The levels of NF kappa B,bFGF, and alpha V beta 3 in A and B groups were significantly lower than those of NS group (P< 0.01) (0.098+/-0.02, 0.156+/-0.02, 0.329+/-0.01; 1.106+/-0.33, 0.412+/-0.01, 1.591+/-0.13; 0.138+/-0.03, 0.114+/-0.02, 0.322+/-0.01). And the level of alpha V beta 3 in C group were significantly reduced compared with those of NS group (P< 0.01) (0.119+/-0.02, 0.322+/-0.01), while the levels of NF kappa B and bFGF had no change between the two groups (P>0.05). The VA/A of the five groups of A, B, C, D, and E were significantly lower than that of NS group (P< 0.05) (26.10+/-13.71, 34.12+/-4.85, 25.50+/-11.41,14.32+/-3.11, 24.36+/-4.95, 66.62+/-17.64), and that of D group were significantly reduced compared with A group (P< 0.05) (14.32+/-3.11, 26.10+/-13.71), but there were no significant difference among the four groups of A, B, C, and E (P >0.05).
Conclusions: NF kappa B can stimulate the angiogenesis of ovarian carcinoma with bFGF by upregulating the expression of alpha V beta 3, bFGF, which can be used as a marker of angiogenesis and a therapy target molecule of ovarian carcinoma.