Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, San Francisco, 505 Parnassus Avenue, San Francisco, California 94143-0132, USA.
Endocrinology. 2010 Mar;151(3):1341-55. doi: 10.1210/en.2009-0923. Epub 2010 Jan 12.
Intrinsic abnormalities in transplanted eutopic endometrium are believed to contribute to the pathogenesis of pelvic endometriosis. Herein we investigated transcriptomic differences in human endometrial stromal fibroblasts (hESFs) from women with (hESF(endo)) vs. without (hESF(nonendo)) endometriosis, in response to activation of the protein kinase A (PKA) pathway with 8-bromoadenosine-cAMP (8-Br-cAMP). hESF(nonendo) (n = 4) and hESF(endo) (n = 4) were isolated from eutopic endometrium and treated +/- 0.5 mm 8-Br-cAMP for 96 h. Purified total RNA was subjected to microarray analysis using the whole-genome Gene 1.0 ST Affymetrix platform. A total of 691 genes were regulated in cAMP-treated hESF(nonendo) vs. 158 genes in hESF(endo), suggesting a blunted response to cAMP/PKA pathway activation in women with disease. Real-time PCR and ELISA validated the decreased expression of decidualization markers in hESF(endo) compared with hESF(nonendo). In the absence of disease, 8-Br-cAMP down-regulated progression through the cell cycle via a decrease in cyclin D1, cyclin-dependent kinase 6, and cell division cycle 2 and an increase in cyclin-dependent kinase inhibitor 1A. However, cell cycle components in hESF(endo) were not responsive to 8-Br-cAMP, resulting in persistence of a proliferative phenotype. hESF(endo) treated with 8-Br-cAMP exhibited altered expression of immune response, extracellular matrix, cytoskeleton, and apoptosis genes. Changes in phosphodiesterase expression and activity were not different among experimental groups. These data support that eutopic hESF(endo) with increased proliferative potential can seed the pelvic cavity via retrograde menstruation and promote establishment, survival, and proliferation of endometriosis lesions, independent of hydrolysis of cAMP and likely due to an inherent abnormality in the PKA pathway.
人们认为，移植的在位子宫内膜中的内在异常有助于盆腔子宫内膜异位症的发病机制。在此，我们研究了存在（hESF（endo））与不存在（hESF（nonendo））子宫内膜异位症的女性子宫内膜基质成纤维细胞（hESF）对蛋白激酶 A（PKA）途径激活的转录组差异，该途径用 8-溴环磷酸腺苷（8-Br-cAMP）激活。从在位子宫内膜中分离 hESF（nonendo）（n = 4）和 hESF（endo）（n = 4），并用 +/- 0.5mm 8-Br-cAMP 处理 96 小时。用全基因组 Gene 1.0 ST Affymetrix 平台对纯化的总 RNA 进行微阵列分析。在 cAMP 处理的 hESF（nonendo）中，有 691 个基因受到调节，而 hESF（endo）中则有 158 个基因受到调节，这表明患有疾病的女性对 cAMP/PKA 途径激活的反应减弱。实时 PCR 和 ELISA 验证了 hESF（endo）中蜕膜化标记物的表达减少与 hESF（nonendo）相比。在没有疾病的情况下，8-Br-cAMP 通过降低 cyclin D1、细胞周期蛋白依赖性激酶 6 和细胞分裂周期 2 的表达和增加细胞周期蛋白依赖性激酶抑制剂 1A，下调细胞周期的进展。然而，hESF（endo）中的细胞周期成分对 8-Br-cAMP 没有反应，导致增殖表型持续存在。用 8-Br-cAMP 处理的 hESF（endo）表现出免疫反应、细胞外基质、细胞骨架和细胞凋亡基因的表达改变。实验组之间磷酸二酯酶表达和活性的变化没有差异。这些数据支持在位 hESF（endo）具有增加的增殖潜能，可以通过逆行月经播种盆腔，并促进子宫内膜异位症病变的建立、存活和增殖，这与 cAMP 的水解无关，可能是由于 PKA 途径的固有异常所致。
