LPA receptor 2 mediates LPA-induced endometrial cancer invasion

Joanie Mayer Hope, Feng qiang Wang, Jill S. Whyte, Edgardo V. Ariztia, Walid Abdalla, Kara Long, David A. Fishman

Research output: Contribution to journalArticle

Abstract

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 μM). 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.

Original languageEnglish (US)
Pages (from-to)215-223
Number of pages9
JournalGynecologic Oncology
Volume112
Issue number1
DOIs
StatePublished - Jan 2009
Externally publishedYes

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Lysophosphatidic Acid Receptors
Endometrial Neoplasms
Matrix Metalloproteinase 7
Gelatin
Conditioned Culture Medium
Small Interfering RNA
lysophosphatidic acid
Cell-Matrix Junctions
Messenger RNA
Collagen Type I
Matrix Metalloproteinases
Ovarian Neoplasms
Transfection
Culture Media
Tail
Cell Culture Techniques

Keywords

  • Endometrial cancer invasion
  • HEC1A
  • LPA receptor 2 (LPA2)
  • Lysophosphatidic acid (LPA)
  • Matrix metalloproteinase-7 (MMP-7)
  • siRNA

ASJC Scopus subject areas

  • Obstetrics and Gynecology
  • Oncology

Cite this

Hope, J. M., Wang, F. Q., Whyte, J. S., Ariztia, E. V., Abdalla, W., Long, K., & Fishman, D. A. (2009). LPA receptor 2 mediates LPA-induced endometrial cancer invasion. Gynecologic Oncology, 112(1), 215-223. https://doi.org/10.1016/j.ygyno.2008.09.019

LPA receptor 2 mediates LPA-induced endometrial cancer invasion. / Hope, Joanie Mayer; Wang, Feng qiang; Whyte, Jill S.; Ariztia, Edgardo V.; Abdalla, Walid; Long, Kara; Fishman, David A.

In: Gynecologic Oncology, Vol. 112, No. 1, 01.2009, p. 215-223.

Research output: Contribution to journalArticle

Hope, JM, Wang, FQ, Whyte, JS, Ariztia, EV, Abdalla, W, Long, K & Fishman, DA 2009, 'LPA receptor 2 mediates LPA-induced endometrial cancer invasion', Gynecologic Oncology, vol. 112, no. 1, pp. 215-223. https://doi.org/10.1016/j.ygyno.2008.09.019
Hope JM, Wang FQ, Whyte JS, Ariztia EV, Abdalla W, Long K et al. LPA receptor 2 mediates LPA-induced endometrial cancer invasion. Gynecologic Oncology. 2009 Jan;112(1):215-223. https://doi.org/10.1016/j.ygyno.2008.09.019
Hope, Joanie Mayer ; Wang, Feng qiang ; Whyte, Jill S. ; Ariztia, Edgardo V. ; Abdalla, Walid ; Long, Kara ; Fishman, David A. / LPA receptor 2 mediates LPA-induced endometrial cancer invasion. In: Gynecologic Oncology. 2009 ; Vol. 112, No. 1. pp. 215-223.
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abstract = "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 μM). 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.",
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AU - Hope, Joanie Mayer

AU - Wang, Feng qiang

AU - Whyte, Jill S.

AU - Ariztia, Edgardo V.

AU - Abdalla, Walid

AU - Long, Kara

AU - Fishman, David A.

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AB - 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 μM). 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.

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