Yoshihiko Ohkawa

Inhibition of 12-O-tetradecanoylphorbol-13-acetate-enhanced transformation in vitro by radical scavengers.

The inhibitory effects of some scavengers of oxygen radicals were studied, using a two-stage transformation assay system in vitro 3-methylcholanthrene (3-MC)-initiation and 12-O-tetradecanoylphorbol-13-acetate-(TPA)-promotion in Balb 3T3 cells. Mannitol, a scavenger for hydroxyl radicals, strongly inhibited the TPA-enhanced transformation in a dose-dependent manner. Superoxide dismutase (SOD) and catalase, which are specific scavengers for O2-. and H2O2, respectively, also inhibited the transformation. These results suggest that oxygen radicals may play an important role in the TPA-enhanced transformation in Balb 3T3 cells.

Retinoic acid and butylated hydroxyanisole inhibit promoter-enhanced transformation in vitro

The inhibitory effects of some antipromoters were studied using a two-stage transformation assay system in vitro with 3-methylcholanthrene (3-MC)-initiation and 12-O-tetradecanoylphorbol-13-acetate (TPA)-promotion in BALB 3T3 cells. Butylated hydroxyanisole (BHA), a phenolic antioxidant, inhibited TPA-enhanced transformation in a dose-dependent manner, but butylated hydroxytoluene (BHT) did not. Among the three antipromoters tested, retinoic acid (RA) was the most effective inhibitor.

A rapid simple screening method for skin tumor promoters using mouse peritoneal macrophages in vitro

The enhancing effects in vitro of a potent skin-tumor promoter, phorbol myristate acetate (PMA), and its derivatives on nitroblue tetrazolium (NBT) reduction, phagocytosis and cell spreading of mouse peritoneal macrophages were compared. The enhancing effects on these markers of macrophage function, especially NBT reduction, were found to correlate well with the skin-tumor promoting activities of the phorbol esters. Another potent promoter, teleocidin, strongly enhanced NBT reduction, although teleocidin is structurally different from phorbol esters. Other tumor promoters, namely saccharin, phenobarbital, cantharidin and Tween 60 did not enhance NBT reduction by macrophages. These data indicate that this test system is appropriate for screening of PMA-type tumor promoters such as phorbol esters and teleocidin.

Induction of nitroblue tetrazolium reduction in mouse peritoneal macrophages by tumor promoters and inhibition of the induced nitroblue tetrazolium reduction by some inhibitors

Two polyacetates, aplysiatoxin and debromoaplysiatoxin, as well as 12-O-tetradecanoylphorbol-13-acetate (TPA), mezerein and teleocidin enhance nitroblue tetrazolium (NBT) reduction in mouse peritoneal macrophages in vitro. The ED50 values for NBT reduction of these 5 TPA-type tumor promoters were 4.2 ng/ml for TPA, 36 ng/ml for mezerein, 0.53 ng/ml for teleocidin, 1.5 ng/ml for aplysiatoxin and 108 ng/ml for debromoaplysiatoxin. The NBT reduction induced by the 5 tumor promoters is inhibited by 2 inhibitors of tumor promotion, retinoic acid and dibromoacetophenone. The possibility that tumor promotion by TPA-type tumor promoters involves similar mechanisms such as superoxide anion radicals release in cell membranes is discussed.

Inhibition of 12-O-tetradecanoylphorbol-13-acetate-induced nitroblue tetrazolium reduction in mouse peritoneal macrophages by various tumor promotion inhibitors

The effects of various inhibitors on 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced reduction of nitroblue tetrazolium (NBT) in mouse peritoneal macrophages were investigated. The reduction was inhibited by phospholipase A2 inhibitors, such as dibromoacetophenone, the lipoxygenase inhibitor nordihydroguaiaretic acid, an NADH-dehydrogenase inhibitor, the microfilament inhibitor cytochalasin B, oxygen radical scavengers such as superoxide dismutase, antioxidants such as butyl hydroxyanisole and non-specific inhibitors such as retinoic acid. The reduction was not affected by the cyclooxygenase inhibitor indomethacin or the H2O2 scavenger catalase.