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Lysophosphatidic acid signaling through LPA receptor subtype 1 induces colony scattering of gastrointestinal cancer cells

  • Kum-Joo Shin
  • You Lim Kim
  • Sukmook Lee
  • Dong-kyu Kim
  • Curie Ahn
  • Junho Chung
  • Jae Young Seong
  • Jong-Ik HwangEmail author
Original Paper

Abstract

Purpose

Lysophosphatidic acid (LPA) is a multifunctional lipid mediator involved in triggering tumor cell invasion and metastasis, as well as malignant cell growth. LPA is also known to modulate the colony scattering of epithelial cancers, which is a prerequisite for cell invasion. However, the underlying details of how this is accomplished are not clear. Here we have investigated the roles of specific LPA receptor subtypes in cell scattering.

Methods

Gastrointestinal carcinoma cell lines were examined for cell scattering activity in response to LPA, and the expression of LPA receptor subtypes was determined by RT-PCR. The effect of down regulation of each LPA receptor in DLD1 cells was determined using a shRNA-lentivirus system. In addition, the effect of overexpression of LPA receptors on cell scattering was investigated using lentivirus expression constructs.

Results

The colonies of AGS and DLD1, but not MKN74, cells were dispersed in response to LPA. RT-PCR analysis revealed that the mRNAs of LPA1, LPA2, and LPA3 were present in AGS and DLD1 cells, but only LPA2 mRNA was detected in MKN74 cells. In DLD1 cells, the scattering activity induced by LPA was partially blocked by pretreatment with PP2 and PD98059, inhibitors of src kinase and MEK, respectively. LPA1 knockdown with shRNA decreased the degree of cell scattering induced by LPA. Knockdown of LPA2 or LPA3 had no effect on LPA-induced scattering. In addition, overexpression of LPA1 in DLD1 cells slightly decreased the response time of LPA-induced cell scattering. On the contrary, MKN74 cells expressing exogenous LPA1 did not respond to LPA by scattering.

Conclusion

These results demonstrate that LPA1 mediates LPA-stimulated cell scattering of gastrointestinal carcinomas, but that activation of other intracellular pathways, besides those contributing to ERK phosphorylation, is also necessary for cell scattering in response to LPA.

Keywords

Lysophosphatidic acid Gastrointestinal carcinomas shRNA Colony scattering Lentivirus 

Abbreviations

LPA

Lysophosphatidic acid

RT-PCR

Reverse transcription-polymerase chain reaction

shRNA

Short hairpin ribonucleic acid

Notes

Acknowledgments

This work was supported by the Korea Research Foundation Grant funded by the Korean Government (MOEHRD)(KRF- 2007-331-C00232) and by the Korea University Grants.

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Kum-Joo Shin
    • 1
  • You Lim Kim
    • 2
  • Sukmook Lee
    • 3
  • Dong-kyu Kim
    • 2
  • Curie Ahn
    • 3
  • Junho Chung
    • 1
  • Jae Young Seong
    • 2
  • Jong-Ik Hwang
    • 2
    Email author
  1. 1.Department of Biochemistry, College of Medicine and Cancer Research InstituteSeoul National UniversitySeoulRepublic of Korea
  2. 2.Graduate School of MedicineKorea UniversitySeoulRepublic of Korea
  3. 3.Clinical Research Institute, Transplantation Research InstituteSeoul National UniversitySeoulRepublic of Korea

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