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 language | English (US) |
|---|---|
| Pages (from-to) | 215-223 |
| Number of pages | 9 |
| Journal | Gynecologic Oncology |
| Volume | 112 |
| Issue number | 1 |
| DOIs | |
| State | Published - Jan 2009 |
| Externally published | Yes |
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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
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 journal › Article
}
TY - JOUR
T1 - LPA receptor 2 mediates LPA-induced endometrial cancer invasion
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.
PY - 2009/1
Y1 - 2009/1
N2 - 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.
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.
KW - Endometrial cancer invasion
KW - HEC1A
KW - LPA receptor 2 (LPA2)
KW - Lysophosphatidic acid (LPA)
KW - Matrix metalloproteinase-7 (MMP-7)
KW - siRNA
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UR - http://www.scopus.com/inward/citedby.url?scp=57649101969&partnerID=8YFLogxK
U2 - 10.1016/j.ygyno.2008.09.019
DO - 10.1016/j.ygyno.2008.09.019
M3 - Article
VL - 112
SP - 215
EP - 223
JO - Gynecologic Oncology
JF - Gynecologic Oncology
SN - 0090-8258
IS - 1
ER -