Rutanen E M, Koistinen R, Wahlström T, Bohn H, Ranta T, Seppälä M
Endocrinology. 1985 Apr;116(4):1304-9. doi: 10.1210/endo-116-4-1304.
The synthesis and secretion of placental protein 12 (PP12) were studied in tissue culture using explants of decidua, amnion, chorion, and placenta from seven full term pregnancies. The total amounts of PP12 in media and tissues were measured by RIA, and new protein synthesis and secretion by decidual explants were demonstrated by the incorporation of [35S]methionine into PP12 after 20 h of incubation with 12.5 microCi/ml [35S]methionine. Cycloheximide was used to study the effect of a protein synthesis inhibitor on the secretion of PP12 by decidua. Significantly more PP12 (P less than 0.001) was released into the medium from decidual explants than from chorion and amnion explants throughout the experimental period of 24 h. When incubated under identical conditions, placental explants released no detectable PP12. In decidual tissues and their culture media, the total amount of PP12 was 127.4% higher after incubation than before incubation (P less than 0.001). No increase was found when chorion and amnion were cultured. The addition of cycloheximide into cultures decreased the total amount of PP12 in the decidua and in its culture medium by more than 50%, indicating that one part of PP12 in decidua was performed and another part was newly synthesized. Decidual explants incorporated [35S]methionine into immunoprecipitable PP12 indicating new PP12 synthesis. In gel filtration, 77% of decidual [35S] PP12 eluted in the same position as purified PP12. In sodium dodecyl sulfate polyacrylamide gel electrophoresis, the migration mobility of [35S]PP12 was identical with that of purified PP12. Our results clearly demonstrate that PP12 is a decidual rather than a placental protein.
利用来自7例足月妊娠的蜕膜、羊膜、绒毛膜和胎盘组织块进行组织培养,研究胎盘蛋白12(PP12)的合成与分泌。采用放射免疫分析法(RIA)测定培养基和组织中PP12的总量,在含有12.5微居里/毫升[35S]甲硫氨酸的培养基中孵育20小时后,通过将[35S]甲硫氨酸掺入PP12来证明蜕膜组织块有新的蛋白质合成与分泌。使用环己酰亚胺研究蛋白质合成抑制剂对蜕膜分泌PP12的影响。在整个24小时的实验期间,与绒毛膜和羊膜组织块相比,从蜕膜组织块释放到培养基中的PP12明显更多(P<0.001)。在相同条件下孵育时,胎盘组织块未释放出可检测到的PP12。在蜕膜组织及其培养基中,孵育后PP12的总量比孵育前高127.4%(P<0.001)。培养绒毛膜和羊膜时未发现增加。向培养物中加入环己酰亚胺使蜕膜及其培养基中PP12的总量减少了50%以上,这表明蜕膜中的PP12一部分是已存在的,另一部分是新合成的。蜕膜组织块将[35S]甲硫氨酸掺入可免疫沉淀的PP12中,表明有新的PP12合成。在凝胶过滤中,77%的蜕膜[35S]PP12与纯化的PP12在相同位置洗脱。在十二烷基硫酸钠聚丙烯酰胺凝胶电泳中,[35S]PP12的迁移率与纯化的PP12相同。我们的结果清楚地表明,PP12是一种蜕膜蛋白而非胎盘蛋白。