Lysophosphatidic acid (LPA) effects on endometrial carcinoma in vitro proliferation, invasion, and matrix metalloproteinase activity

Gynecol Oncol. 2010 Apr;117(1):88-95. doi: 10.1016/j.ygyno.2009.12.012. Epub 2010 Jan 6.

Abstract

Objectives: 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.

Methods: 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.

Results: 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.

Conclusions: 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.

MeSH terms

  • Carcinoma, Endometrioid / enzymology*
  • Carcinoma, Endometrioid / pathology*
  • Cell Growth Processes / drug effects
  • Cell Line, Tumor
  • Endometrial Neoplasms / enzymology*
  • Endometrial Neoplasms / pathology*
  • Enzyme Activation / drug effects
  • Female
  • Fibrosarcoma / enzymology
  • Fibrosarcoma / pathology
  • Humans
  • Lysophospholipids / pharmacology*
  • Matrix Metalloproteinase 2 / biosynthesis
  • Matrix Metalloproteinase 2 / genetics
  • Matrix Metalloproteinase 2 / metabolism
  • Matrix Metalloproteinase 7 / biosynthesis
  • Matrix Metalloproteinase 7 / genetics
  • Matrix Metalloproteinase 7 / metabolism
  • Matrix Metalloproteinases / biosynthesis
  • Matrix Metalloproteinases / genetics
  • Matrix Metalloproteinases / metabolism*
  • Neoplasm Invasiveness
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics

Substances

  • Lysophospholipids
  • RNA, Messenger
  • Matrix Metalloproteinases
  • Matrix Metalloproteinase 7
  • Matrix Metalloproteinase 2
  • lysophosphatidic acid