Wegner C C, Carson D D
Department of Biochemistry and Molecular Biology, University of Texas M. D. Anderson Cancer Center, Houston 77030.
Endocrinology. 1992 Dec;131(6):2565-72. doi: 10.1210/endo.131.6.1446600.
While uterine stromal cells (USC) appear to modify the function of uterine epithelial cells (UEC) under certain conditions in vivo, relatively little is known about the effect of epithelial cells on stromal cell differentiation and function. To determine if UEC modulate USC function in vitro, highly enriched (> 95%) cultures of polarized UEC were first cultured on Matrigel-coated filters in serum-free medium until confluent, then cocultured with USC for up to 120 h. Subsequently, while maintaining both cell types in physically separate compartments, filters containing UEC were removed, and USC phenotypic markers assayed. Coculture with UEC did not affect the expression of two markers of USC differentiation (desmin and laminin), USC DNA content, [35S]methionine uptake, or total protein synthesis or secretion. However, coculture of USC with UEC or medium conditioned by UEC induced the secretion of a 30-kilodalton protein (p30) from USC as early as 24 h of coculture and through 120 h of coculture. In addition, secretion of a 60-kilodalton protein by USC was frequently observed in response to coculture with UEC. Neither the hormonal stage from which uterine cells were recovered, nor the addition of exogenous progesterone or estradiol modulated UEC-induced p30 secretion. Several purified growth factors (transforming growth factor-beta, epidermal growth factor, interleukin-1 alpha, and fibroblast growth factor) added to the serum-free culture medium failed to induce p30 secretion by USC. The p30-inducing activity in UEC-conditioned medium could not be abolished by either heat or trypsin treatment, suggesting that it is not a protein. Purified prostaglandin E2 or F2 alpha or platelet-activating factor did not induce p30 secretion by isolated USC. Of several epithelial and fibroblastic cell lines tested, UEC and a human uterine adenocarcinoma cell line (RL95-2) were the most effective in inducing p30 secretion by USC. Moreover, UEC also were able to modulate protein secretion by nonuterine murine fibroblast cell lines. Collectively, these data demonstrate that UEC can modulate USC function in vitro via a soluble factor(s).
虽然在体内某些条件下子宫基质细胞（USC）似乎会改变子宫上皮细胞（UEC）的功能，但关于上皮细胞对基质细胞分化和功能的影响却知之甚少。为了确定UEC在体外是否调节USC功能，首先将高度富集（>95%）的极化UEC培养物在无血清培养基中培养在基质胶包被的滤器上，直至汇合，然后与USC共培养长达120小时。随后，在将两种细胞类型保持在物理上分开的隔室的同时，移除含有UEC的滤器，并检测USC表型标记物。与UEC共培养不影响USC分化的两个标记物（结蛋白和层粘连蛋白）的表达、USC DNA含量、[35S]甲硫氨酸摄取或总蛋白合成或分泌。然而，USC与UEC或UEC条件培养基共培养早在共培养24小时就诱导USC分泌一种30千道尔顿的蛋白质（p30），并持续到共培养120小时。此外，经常观察到USC对与UEC共培养的反应是分泌一种60千道尔顿的蛋白质。回收子宫细胞的激素阶段，以及添加外源性孕酮或雌二醇均未调节UEC诱导的p30分泌。添加到无血清培养基中的几种纯化生长因子（转化生长因子-β、表皮生长因子、白细胞介素-1α和成纤维细胞生长因子）未能诱导USC分泌p30。UEC条件培养基中的p30诱导活性不能通过加热或胰蛋白酶处理消除，这表明它不是一种蛋白质。纯化的前列腺素E2或F2α或血小板活化因子不能诱导分离的USC分泌p30。在测试的几种上皮和成纤维细胞系中，UEC和人子宫腺癌细胞系（RL95-2）在诱导USC分泌p30方面最有效。此外，UEC还能够调节非子宫小鼠成纤维细胞系的蛋白质分泌。总体而言，这些数据表明UEC可以通过一种或多种可溶性因子在体外调节USC功能。
