LPA receptor 2 mediates LPA-induced endometrial cancer invasion
LPA receptor 2 mediates LPA-induced endometrial cancer invasion
复制标题
DOI：
10.1016/j.ygyno.2008.09.019
复制
发表时间：
2009-01-01
影响因子：
4.7
通讯作者：
Fishman, David A.
中科院分区：
文献类型：
--
作者：
Hope, Joanie Mayer;Wang, Feng-qiang;Fishman, David A.
目标。我们先前已经证明溶血磷脂酸(LPA)促进卵巢癌转移级联反应。在这项研究中，我们评估了LPA在子宫内膜癌侵袭中的作用。使用预先设计的针对LPA受体2(LPA2)和人基质金属蛋白酶-7(MMP-7)的siRNA双链完成瞬时mRNA敲除。逆转录聚合酶链式反应检测LPA受体和基质金属蛋白酶-7的表达。用鼠尾胶原I型包被Boyden小室进行体外侵袭分析。明胶酶谱检测细胞培养条件培养液中的基质金属蛋白酶活性。在LPA2基因敲除后，细胞-细胞和细胞-基质附着也被评估，以进一步阐明LPA2级联反应。LPA在生理浓度(0.1-1mM)时可增加HEC1A细胞的侵袭力。在四种主要的LPA受体中，LPA2主要由HEC1A细胞表达。瞬时转染LPA2siRNA可使HEC1A细胞LPA2mRNA表达降低93%(P<；0.01)。沉默LPA2可消除LPA刺激的侵袭增加(P<；0.05)，减少LPA诱导的基质金属蛋白酶-7的分泌/激活，而不显著影响细胞与细胞或细胞与基质的黏附。与阴性对照相比，沉默基质金属蛋白酶-7降低了细胞的整体侵袭能力，但不能消除LPA对HEC1A细胞的侵袭作用(P<；0.05)。明胶酶谱证实，LPA2和MMP7基因敲除可降低HEC1A条件培养液中MMP7的活性。LPA2介导LPA刺激的HEC1A侵袭和随后激活的基质金属蛋白酶-7。(C)2008 Elsevier Inc.保留所有权利。
Objective. We have previously shown that lysophosphatidic acid (LPA) promotes the ovarian cancer metastatic cascade. In this study, we evaluated the role of LPA on endometrial cancer invasion.Methods. Transient mRNA knockdown was accomplished using pre-designed siRNA duplexes against LPA receptor 2 (LPA2) and human matrix metalloproteinase-7 (MMP-7). RT-PCR was used to characterize LPA receptor and MMP-7 expression. Analysis of in vitro invasion was performed with rat-tail collagen type I coated Boyden chambers. Gelatin zymography was used to evaluate the MMP activity in cell culture conditioned media. Cell-cell and cell-matrix attachment was also assessed upon LPA2 knockdown to further illuminate the LPA2 cascade.Results. LPA increases HEC1A cellular invasion at physiologic concentrations (0.1-1 mu M). Of the four principle LPA receptors, LPA2 is predominantly expressed by HEC1A cells. Transient transfection of LPA2 siRNA reduced LPA2 mRNA expression in HEC1A cells by 93% (P < 0.01). Silencing LPA2 eliminated the LPA-stimulated increase in invasion (P < 0.05) and reduced LPA-induced MMP-7 secretion/activation, without significantly affecting cell-cell or cell-matrix adhesion. Silencing MMP-7 reduced overall invasion but did not eliminate LPA's pro-invasive effect on HEC1A cells, as compared to negative control (P < 0.05). Gelatin zymography confirmed that LPA2 and MMP-7 knockdown reduced MMP-7 activation in HEC1A conditioned media.Conclusion. LPA2 mediates LPA-stimulated HEC1A invasion and the subsequent activation of MMP-7. (c) 2008 Elsevier Inc. All rights reserved.