Haaparanta T, Halpert J, Glaumann H, Gustafsson J A
Cancer Res. 1983 Nov;43(11):5131-7.
Treatment with beta-naphthoflavone (BNF) was found to induce 7-ethoxyresorufin O-deethylase and aryl hydrocarbon hydroxylase activities about 500-fold in the microsomal fraction of the rat ventral prostate but had no effect on aminopyrine N-demethylase or reduced nicotinamide adenine dinucleotide phosphate:cytochrome c reductase activities. Phenobarbital (PB) treatment did not alter any of these enzyme activities. Antibodies raised in rabbits against rat liver cytochrome P-450 reductase (P-450 reductase) and against P-450 BNF-B2 and P-450 PB-B2, the major forms of P-450 isolated from liver microsomes of BNF- and PB-treated rats, respectively, were used to characterize the P-450-dependent monooxygenase system in the rat ventral prostate. Anti-P-450 reductase immunoglobulin G inhibited reduced nicotinamide adenine dinucleotide phosphate:cytochrome c reductase activity in prostatic microsomes, and anti-P-450 BNF-B2 but not anti-P-450 PB-B2 immunoglobulin G inhibited the BNF-induced prostatic microsomal 7-ethoxyresorufin O-deethylase and aryl hydrocarbon hydroxylase activities. A highly sensitive immunoblotting method was used to quantitate P-450 BNF-B2, P-450 PB-B2, and P-450 reductase in prostatic microsomes. Using this technique, prostatic P-450 reductase with a molecular weight corresponding to that of purified liver P-450 reductase was detected at a level of 0.02 nmol/mg of microsomal protein. In the liver, the same enzyme amounts to 0.2 nmol/mg of microsomal protein. P-450 BNF-B2 was not detected in prostatic microsomes from control or PB-treated rats, whereas a protein band with a molecular weight corresponding to that of purified liver P-450 BNF-B2 was found in prostatic microsomes from BNF-treated rats at a level of 0.05 nmol P-450 per mg microsomal protein. P-450 PB-B2 was not detected in prostatic microsomes from either control, PB-treated, or BNF-treated animals.
研究发现，用β-萘黄酮（BNF）处理可使大鼠腹侧前列腺微粒体部分的7-乙氧基试卤灵O-脱乙基酶和芳烃羟化酶活性诱导增加约500倍，但对氨基比林N-脱甲基酶或还原型烟酰胺腺嘌呤二核苷酸磷酸：细胞色素c还原酶活性没有影响。苯巴比妥（PB）处理未改变这些酶活性中的任何一种。用兔抗大鼠肝细胞色素P-450还原酶（P-450还原酶）以及分别针对从BNF和PB处理大鼠的肝微粒体中分离出的主要P-450形式P-450 BNF-B2和P-450 PB-B2产生的抗体，来表征大鼠腹侧前列腺中依赖P-450的单加氧酶系统。抗P-450还原酶免疫球蛋白G抑制前列腺微粒体中还原型烟酰胺腺嘌呤二核苷酸磷酸：细胞色素c还原酶活性，而抗P-450 BNF-B2免疫球蛋白G而非抗P-450 PB-B2免疫球蛋白G抑制BNF诱导的前列腺微粒体7-乙氧基试卤灵O-脱乙基酶和芳烃羟化酶活性。采用一种高灵敏度免疫印迹法对前列腺微粒体中的P-450 BNF-B2、P-450 PB-B2和P-450还原酶进行定量。利用该技术，检测到前列腺中分子量与纯化的肝P-450还原酶相对应的P-450还原酶，含量为0.02 nmol/mg微粒体蛋白。在肝脏中，该酶含量为0.2 nmol/mg微粒体蛋白。在对照或PB处理大鼠的前列腺微粒体中未检测到P-450 BNF-B2，而在BNF处理大鼠的前列腺微粒体中发现一条分子量与纯化的肝P-450 BNF-B2相对应的蛋白条带，含量为0.05 nmol P-450/mg微粒体蛋白。在对照、PB处理或BNF处理动物的前列腺微粒体中均未检测到P-450 PB-B2。
