Department of Obstetrics, Gynecology and Reproductive Sciences, Mount Sinai School of Medicine, New York, NY 10029, USA.
Gynecol Oncol. 2010 Apr;117(1):88-95. doi: 10.1016/j.ygyno.2009.12.012. Epub 2010 Jan 6.
Lysophosphatidic acid (LPA) has potent growth-regulatory effect in many cell types and has been linked to the in vivo tumor growth and metastasis in several malignancies. The goal of this study was to assess the regulation of (EC) microenvironment by LPA through the examination of its effect on cell proliferation, migration, invasion, uPA activity, and matrix metalloproteinase (MMP) secretion/activation.
All experiments were performed in vitro using an EC cell line, HEC-1A. Cell proliferation was determined using the Promega MTS proliferation assay following 48 h of exposures to different concentrations of LPA (0.1, 1.0 and 10.0 microM). Cell invasion was assessed using a modified Boyden chamber assay with collagen I coated on the membrane. HEC-1A motility was examined by Boyden chamber migration assay as well as the scratch wound closure assay on type I collagen. MMP secretion/activation in HEC-1A conditioned medium was detected by gelatin zymography. MMP-7 mRNA expression was determined using real-time PCR. uPA activity was measured using a coupled colorimetric assay.
LPA, at the concentrations of 0.1 and 1.0 microM, significantly induced the proliferation of HEC-1A cells (p<0.01). At 10 microM, LPA- induced HEC-1A proliferation to a less extent and showed no significant effect on HEC-1A invasion and migration (p>0.05). Gelatin zymogram showed that HEC-1A cells secreted high levels of MMP-7, while MMP-2 and MMP-9 are barely detectable. In addition, LPA significantly enhanced uPA activity in HEC-1A conditioned medium in a concentration-dependent manner.
LPA is a potent modulator of cellular proliferation and invasion for EC cells. It also has the capacity to stimulate the secretion/activity of uPA and MMP-7. Those results suggest that LPA is a bioactive modulator of EC microenvironment and may have a distinct regulation mechanism as observed in epithelial ovarian cancer.
溶血磷脂酸（LPA）在许多细胞类型中具有强大的生长调节作用，并与几种恶性肿瘤的体内肿瘤生长和转移有关。本研究的目的是通过研究其对细胞增殖、迁移、侵袭、uPA 活性和基质金属蛋白酶（MMP）分泌/激活的影响来评估（EC）微环境的调节。
所有实验均在体外使用 EC 细胞系 HEC-1A 进行。细胞增殖通过 Promega MTS 增殖测定法在暴露于不同浓度 LPA（0.1、1.0 和 10.0 microM）48 小时后确定。细胞侵袭通过在膜上涂覆胶原 I 的改良 Boyden 室测定法进行评估。HEC-1A 运动通过 Boyden 室迁移测定法以及 I 型胶原上的划痕伤口闭合测定法进行检查。HEC-1A 条件培养基中的 MMP 分泌/激活通过明胶酶谱法检测。使用实时 PCR 确定 MMP-7 mRNA 表达。使用偶联比色测定法测量 uPA 活性。
0.1 和 1.0 microM 的 LPA 浓度显著诱导 HEC-1A 细胞增殖（p<0.01）。在 10 microM 时，LPA 诱导的 HEC-1A 增殖程度较小，对 HEC-1A 侵袭和迁移没有显著影响（p>0.05）。明胶酶谱显示 HEC-1A 细胞分泌高水平的 MMP-7，而 MMP-2 和 MMP-9 几乎检测不到。此外，LPA 以浓度依赖性方式显著增强 HEC-1A 条件培养基中的 uPA 活性。
LPA 是 EC 细胞增殖和侵袭的有力调节剂。它还具有刺激 uPA 和 MMP-7 分泌/活性的能力。这些结果表明 LPA 是 EC 微环境的生物活性调节剂，并且可能具有与上皮性卵巢癌观察到的不同的调节机制。
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