Rutanen E M, Menabawey M, Isaka K, Bohn H, Chard T, Grudzinskas J G
J Clin Endocrinol Metab. 1986 Sep;63(3):675-9. doi: 10.1210/jcem-63-3-675.
The synthesis and secretion of placental protein 12 (PP12) by early pregnancy decidua and trophoblast were studied in vitro from tissues obtained by curettage during elective termination of pregnancy (weeks 8-14). The tissue explants were incubated in Ham's F-10 medium for a 27-h period, and the PP12 levels in media and tissue homogenates were measured by RIA. De novo synthesis of PP12 was assessed by measuring the incorporation of radioactivity into PP12 after 20 h of incubation of tissues with 20 microCi/ml [35S]methionine. PP12 from the culture medium was immunoprecipitated with anti-PP12(A) antiserum, and the immunoprecipitate was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The initial tissue content of PP12 was 10- to 72-fold higher in decidua than in trophoblast. The total amount of radioimmunoassayable PP12 released into medium by decidual explants during the 27-h incubation period together with that present in the tissues at the end of incubation exceeded the initial tissue content by 242.7 +/- 63.7% (mean +/- SE). Only small amounts of PP12 were detected in media from trophoblast cultures. During the first 7 h of incubation, inclusion of cycloheximide had no effect on PP12 release by decidual explants in three of four experiments. Between 7 and 27 h, the amount of PP12 released by cycloheximide-treated tissues was 20.0 +/- 7% of that released by control tissues (P less than 0.01). Cycloheximide had no effect on PP12 release by trophoblasts. Decidual explants incorporated [35S]methionine into PP12, but trophoblasts did not. In sodium dodecyl sulfate-gel electrophoresis, the newly synthesized PP12 comigrated with the major band of purified PP12 corresponding to mol wt 29,000. These data clearly confirm that PP12 is a protein of decidual rather than trophoblastic origin, and indicate that decidua from early pregnancy has the ability to synthesize it.
对在妊娠早期(8 - 14周)选择性终止妊娠时刮宫获取的组织进行体外研究,观察早孕蜕膜和滋养层对胎盘蛋白12(PP12)的合成与分泌情况。将组织外植体在Ham's F - 10培养基中孵育27小时,采用放射免疫分析法(RIA)测定培养基和组织匀浆中的PP12水平。通过在含20微居里/毫升[35S]甲硫氨酸的培养基中孵育组织20小时后,测量放射性物质掺入PP12的量来评估PP12的从头合成。用抗PP12(A)抗血清对培养基中的PP12进行免疫沉淀,并用十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳分析免疫沉淀物。PP12的初始组织含量在蜕膜中比在滋养层中高10至72倍。在27小时的孵育期内,蜕膜外植体释放到培养基中的可放射免疫测定的PP12总量以及孵育结束时组织中存在的PP12总量比初始组织含量高出242.7±63.7%(平均值±标准误)。在滋养层培养物的培养基中仅检测到少量PP12。在孵育的前7小时,在四个实验中的三个实验中,加入环己酰亚胺对蜕膜外植体释放PP12没有影响。在7至27小时之间,经环己酰亚胺处理的组织释放的PP12量是对照组织释放量的20.0±7%(P<0.01)。环己酰亚胺对滋养层释放PP12没有影响。蜕膜外植体将[35S]甲硫氨酸掺入PP12,但滋养层没有。在十二烷基硫酸钠凝胶电泳中,新合成的PP12与纯化的PP12的主要条带迁移率相同,对应分子量为29,000。这些数据清楚地证实PP12是蜕膜来源而非滋养层来源的蛋白质,并表明早孕蜕膜有能力合成它。