Ovarian cancer cells stimulate uPA gene expression in fibroblastic stromal cells via multiple paracrine and autocrine mechanisms
Ovarian cancer cells stimulate uPA gene expression in fibroblastic stromal cells via multiple paracrine and autocrine mechanisms
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DOI：
10.1016/j.ygyno.2009.06.026
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发表时间：
2009-10-01
影响因子：
4.7
通讯作者：
Casslen, Bertil
中科院分区：
文献类型：
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作者：
Noskova, Vera;Ahmadi, Sara;Casslen, Bertil
目标。UPA mRNA在低分化卵巢肿瘤间质中的表达显著上调。我们假设这种表达是由上皮性肿瘤细胞旁分泌信号诱导的，并建立了卵巢癌微环境的体外模型来研究细胞间的相互作用。ES-2透明细胞癌细胞在组织培养片中生长在双室系统中，成纤维基质LEP细胞包埋在Matrigel中。在与LEP细胞孵育前，使用结合位点定向抗体中和ES-2条件培养液中的可溶性细胞因子。实时荧光定量聚合酶链式反应检测LEP细胞中uPA基因的表达，以及两种细胞中细胞因子的基因表达。与ES-2细胞共培养以及与ES-2 CM共同孵育可诱导LEP细胞uPA mRNA表达增加约一倍。在LEP细胞与CM孵育的短时间内(12h)，抗EGF和bFGF的抗体减少了uPA mRNA的诱导，提示这些细胞因子起旁分泌信号的作用。ES-2细胞中也有EGF和bFGFmRNA表达。在较长时间孵育(24 H)时，抗bFGF、HB-EGF、HGF、IGF-1和IL-1α的抗体降低了uPA mRNA的诱导，提示这些细胞因子在LEP细胞中具有自分泌功能。事实上，在染毒的LEP细胞中，上述5种细胞因子的表达均上调。在卵巢癌细胞中，我们发现两种细胞因子是旁分泌信号，五种细胞因子是自分泌信号，它们诱导间质成纤维细胞uPA mRNA表达上调。了解肿瘤内的串扰是至关重要的，因为它可以提供新的治疗方法。(C)2009 Elsevier Inc.保留所有权利。
Objectives. Expression Of uPA mRNA is massively up-regulated in the stroma of poorly differentiated ovarian tumors. We hypothesized that this expression was induced by paracrine signals from the epithelial tumor cells, and established an in vitro model of ovarian cancer microenvironment to study intercellular cross-talk.Methods. ES-2 clear cell carcinoma cells were grown in tissue culture inserts in a double-chamber system with fibroblastic stromal LEP cells embedded in Matrigel. Binding-site directed antibodies were used to neutralize soluble cytokines in ES-2 conditioned medium (CM) before incubation with LEP cells. Real time PCR measured uPA mRNA in LEP cells, as well as mRNA for cytokines in both cell types.Results. Co-culture with ES-2 cells as well as incubation with ES-2 CM induced uPA mRNA in LEP cells about two-fold. In short time (12 h) incubation of LEP cells with CM, antibodies to EGF and bFGF reduced induction of uPA mRNA, suggesting that these cytokines function as paracrine signals. EGF mRNA and bFGF mRNA were also found in ES-2 cells. At longer incubation (24 h) antibodies to bFGF, HB-EGF, HGF, IGF-1, and IL-1 alpha reduced uPA mRNA induction, suggesting an autocrine function for these cytokines in LEP cells. In fact, expression of the same five cytokines was up-regulated in LEP cells exposed to CM.Conclusion. We identified two cytokines as paracrine signals, and five cytokines as autocrine signals in ovarian cancer cell induced up-regulation of uPA mRNA in stromal fibroblastic cells. It is crucial to understand intra-tumoral cross-talk, since it can offer new therapeutic approaches. (C) 2009 Elsevier Inc. All rights reserved.