Medical Research Council Human Reproductive Sciences Unit, The Queen's Medical Research Institute, The University of Edinburgh, Edinburgh, EH16 4TJ, UK.
BMC Cancer. 2010 Sep 14;10:488. doi: 10.1186/1471-2407-10-488.
An increase in cancer cell invasion and microvascular density is associated with a poorer prognosis for patients with endometrial cancer. In endometrial adenocarcinoma F-prostanoid (FP) receptor expression is elevated, along with its ligand prostaglandin (PG)F2α, where it regulates expression and secretion of a host of growth factors and chemokines involved in tumorigenesis. This study investigates the expression, regulation and role of a disintegrin and metalloproteinase with thrombospondin repeat 1 (ADAMTS1) in endometrial adenocarcinoma cells by PGF2α via the FP receptor.
Human endometrium and adenocarcinoma tissues were obtained in accordance with Lothian Research Ethics Committee guidance with informed patient consent. Expression of ADAMTS1 mRNA and protein in tissues was determined by quantitative RT-PCR analysis and immunohistochemistry. Signal transduction pathways regulating ADAMTS1 expression in Ishikawa cells stably expressing the FP receptor to levels seen in endometrial cancer (FPS cells) were determined by quantitative RT-PCR analysis. In vitro invasion and proliferation assays were performed with FPS cells and human umbilical vein endothelial cells (HUVECs) using conditioned medium (CM) from PGF2α-treated FPS cells from which ADAMTS1 was immunoneutralised and/or recombinant ADAMTS1. The role of endothelial ADAMTS1 in endothelial cell proliferation was confirmed with RNA interference. The data in this study were analysed by T-test or ANOVA.
ADAMTS1 mRNA and protein expression is elevated in endometrial adenocarcinoma tissues compared with normal proliferative phase endometrium and is localised to the glandular and vascular cells. Using FPS cells, we show that PGF2α-FP signalling upregulates ADAMTS1 expression via a calmodulin-NFAT-dependent pathway and this promotes epithelial cell invasion through ECM and inhibits endothelial cell proliferation. Furthermore, we show that CM from FPS cells regulates endothelial cell ADAMTS1 expression in a rapid biphasic manner. Using RNA interference we show that endothelial cell ADAMTS1 also negatively regulates cellular proliferation.
These data demonstrate elevated ADAMTS1 expression in endometrial adenocarcinoma. Furthermore we have highlighted a mechanism whereby FP receptor signalling regulates epithelial cell invasion and endothelial cell function via the PGF2α-FP receptor mediated induction of ADAMTS1.
癌细胞侵袭和微血管密度的增加与子宫内膜癌患者预后不良有关。在子宫内膜腺癌中，F 型前列腺素（FP）受体表达升高，其配体前列腺素（PG）F2α也升高，它调节参与肿瘤发生的一系列生长因子和趋化因子的表达和分泌。本研究通过 FP 受体研究了 PGF2α 对子宫内膜腺癌细胞中解整合素和金属蛋白酶与凝血酶重复 1（ADAMTS1）的表达、调节和作用。
根据洛锡安研究伦理委员会的指导和患者知情同意，获得人子宫内膜和腺癌组织。通过定量 RT-PCR 分析和免疫组织化学测定组织中 ADAMTS1 mRNA 和蛋白的表达。通过定量 RT-PCR 分析，确定稳定表达 FP 受体的 Ishikawa 细胞中调节 ADAMTS1 表达的信号转导途径，其表达水平与子宫内膜癌相似（FPS 细胞）。使用来自经 PGF2α 处理的 FPS 细胞的条件培养基（CM），在 FPS 细胞和人脐静脉内皮细胞（HUVEC）中进行体外侵袭和增殖测定，其中 ADAMTS1 被免疫中和和/或重组 ADAMTS1。使用 RNA 干扰证实内皮 ADAMTS1 在内皮细胞增殖中的作用。本研究中的数据通过 T 检验或 ANOVA 进行分析。
与正常增生期子宫内膜相比，子宫内膜腺癌组织中 ADAMTS1 mRNA 和蛋白表达升高，定位于腺细胞和血管细胞。使用 FPS 细胞，我们表明 PGF2α-FP 信号通过钙调蛋白-NFAT 依赖性途径上调 ADAMTS1 表达，这促进上皮细胞通过 ECM 侵袭并抑制内皮细胞增殖。此外，我们表明 FPS 细胞的 CM 以快速双相方式调节内皮细胞 ADAMTS1 的表达。通过 RNA 干扰，我们表明内皮细胞 ADAMTS1 也负调节细胞增殖。
这些数据表明子宫内膜腺癌中 ADAMTS1 表达升高。此外，我们强调了一种机制，即 FP 受体信号通过 PGF2α-FP 受体介导的 ADAMTS1 诱导调节上皮细胞侵袭和内皮细胞功能。
