Salamonsen L A, Healy D L, Findlay J K
Medical Research Centre, Prince Henry's Hospital, Melbourne, Victoria, Australia.
J Steroid Biochem. 1987 Sep;28(3):285-8. doi: 10.1016/0022-4731(87)91020-x.
Cultured ovine epithelial endometrial cells from oestrogen-treated ovariectomized ewes were treated in vitro with combinations of oestradiol-17 beta (E), progesterone (P) and the P-receptor antagonist RU486 (each 10(-6) to 10(-9) M), in the presence of [35S]methionine. Neither DNA content of dishes nor total protein were increased in treatment compared with control dishes. Incorporation of [35S] into secreted protein was lower from cells treated in vitro with P or E + P (10(-9) M) than from those treated with E (10(-9) M, P less than 0.01). Incorporation of [35S] into cellular protein was decreased by P (10(-9) M, P less than 0.025). SDS-PAGE analysis of secreted proteins enabled measurement of levels of a 46K protein which is secreted maximally following E + P administration in vivo. In vitro, P either alone or with E (each 10(-7) M) increased the abundance of the 46K protein in cell secretions by a factor of 1.5 +/- 0.1 (N = 9) or 1.8 +/- 0.3 (N = 10) respectively (P less than 0.01) compared with controls. The administration of E (10(-7) M) or either or both steroids at 10(-9) M, was without effect. RU486 alone (10(-6) to 10(-8) M) was also without effect but in the presence of E + P or P, blocked the increase in the 46K protein, suggesting this effect is mediated via binding of P to its receptor.
从经雌激素处理的去卵巢母羊中培养的绵羊子宫内膜上皮细胞，在存在[35S]甲硫氨酸的情况下，于体外用17β-雌二醇（E）、孕酮（P）和孕酮受体拮抗剂RU486（浓度均为10(-6)至10(-9) M）的组合进行处理。与对照培养皿相比，处理组培养皿中的DNA含量和总蛋白均未增加。用P或E + P（10(-9) M）体外处理的细胞，其分泌蛋白中[35S]的掺入量低于用E（10(-9) M）处理的细胞（P < 0.01）。P（10(-9) M）使细胞蛋白中[35S]的掺入量减少（P < 0.025）。对分泌蛋白进行SDS-PAGE分析，可以测定一种46K蛋白的水平，该蛋白在体内给予E + P后分泌量最大。在体外，单独的P或与E（各为10(-7) M）一起，与对照相比，分别使细胞分泌物中46K蛋白的丰度增加了1.5 +/- 0.1（N = 9）倍或1.8 +/- 0.3（N = 10）倍（P < 0.01）。给予E（10(-7) M）或10(-9) M的任一或两种类固醇均无作用。单独的RU486（10(-6)至10(-8) M）也无作用，但在存在E + P或P的情况下，可阻断46K蛋白的增加，表明这种作用是通过P与其受体结合介导的。
