溶血磷脂酸（LPA）以尿激酶型纤溶酶原激活物（uPA）依赖的方式促进E-钙黏蛋白胞外结构域脱落和OVCA429细胞侵袭。

Lysophosphatidic acid (LPA) promotes E-cadherin ectodomain shedding and OVCA429 cell invasion in an uPA-dependent manner.

作者信息

Gil Orlando D, Lee Catherine, Ariztia Edgardo V, Wang Feng-Qiang, Smith Phillip J, Hope Joanie Mayer, Fishman David A

机构信息

Department of Obstetrics and Gynecology, New York University School of Medicine, 550 First Avenue, TH-528, New York, NY 10016, USA.

出版信息

Gynecol Oncol. 2008 Feb;108(2):361-9. doi: 10.1016/j.ygyno.2007.10.027.

DOI:10.1016/j.ygyno.2007.10.027

PMID:18073130

Abstract

OBJECTIVES

To evaluate the role of LPA in regulating E-cadherin cell surface expression, adhesion, and invasion in epithelial ovarian carcinoma (EOC) cells.

METHODS

E-cadherin mRNA expression in OVCA429 and IOSE-29 cells was evaluated by real-time RT-PCR. Immunofluorescence and Western blot analysis were performed to determine cell surface expression and shedding of E-cadherin 80-kDa soluble fragment by LPA. Kinetics of LPA-induced uPA activity was followed with a colorimetric enzymatic assay. Invasion assays were performed in a modified Boyden chamber where cells were allowed to migrate to the bottom compartment through a porous filter coated with collagen. Additionally we measured the 80-kDa form from the ascites of women with stage III/IV EOC.

RESULTS

LPA induces E-cadherin shedding of a soluble 80-kDa fragment. We found that this process is mediated by the uPA proteolytic cascade. High levels of soluble E-cadherin were found in the ascites from women with advanced stage EOC. LPA and a soluble recombinant E-cadherin-Fc chimera promotes invasion of OVCA429 cells.

CONCLUSIONS

LPA induces shedding of an 80-kDa E-cadherin-soluble fragment in an uPA-dependent manner and promotes in vitro invasion. High levels of soluble E-cadherin in malignant ascites may also affect ovarian metastasis.

摘要

目的

评估溶血磷脂酸（LPA）在上皮性卵巢癌（EOC）细胞中调节E-钙黏蛋白细胞表面表达、黏附及侵袭的作用。

方法

通过实时逆转录聚合酶链反应（RT-PCR）评估OVCA429和IOSE-29细胞中E-钙黏蛋白mRNA的表达。进行免疫荧光和蛋白质印迹分析，以确定LPA对E-钙黏蛋白80-kDa可溶性片段的细胞表面表达及脱落情况。采用比色酶法追踪LPA诱导的尿激酶型纤溶酶原激活物（uPA）活性动力学。在改良的博伊登小室中进行侵袭实验，使细胞通过包被有胶原蛋白的多孔滤膜迁移至下室。此外，我们还检测了III/IV期EOC女性患者腹水中的80-kDa形式。