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Digestive Diseases and Sciences

, Volume 59, Issue 7, pp 1428–1435 | Cite as

Piezo1 Is as a Novel Trefoil Factor Family 1 Binding Protein that Promotes Gastric Cancer Cell Mobility In Vitro

  • Xiao-Ning Yang
  • Ya-Pi Lu
  • Jing-Jing Liu
  • Jian-Kun Huang
  • Yun-Peng Liu
  • Chuan-Xing Xiao
  • Amarsanaa Jazag
  • Jian-Lin RenEmail author
  • Bayasi GulengEmail author
Original Article

Abstract

Background

Trefoil factor family 1 (TFF1) is a member of the TFF-domain peptide family involved in epithelial restitution and cell motility. Recently, we screened Piezo1 as a candidate TFF1-binding protein.

Aim

We aimed to confirm Piezo1 as a novel TFF1 binding protein and to assess the role of this interaction in mediating gastric cancer cell mobility.

Methods

This interaction was confirmed by co-immunoprecipitation and co-localisation of TFF1 and Piezo1 in GES-1 cells. We used stable RNA interference to knockdown Piezo1 protein expression and restored the expression of TFF1 in the gastric cancer cell lines SGC-7901 and BGC-823. Cell motility was evaluated using invasion assay and migration assay in vitro. The expression levels of the integrin subunits β1, β5, α1 as well as the expression of β-catenin and E-cadherin were detected by Western blot.

Results

We demonstrate that TFF1, but not TFF2 or TFF3, bind to and co-localize with Piezo1 in the cytoplasm in vitro. TFF1 interacts with the C-terminal portion of the Piezo1 protein. Wound healing and trans-well assays demonstrated that the restored expression of TFF1 promoted cell mobility in gastric cancer cells, and this effect was attenuated by the knockdown of Piezo1. Western blots demonstrated the decreased expression of integrin β1 in Piezo1-knockdown cells.

Conclusions

Our data demonstrate that Piezo1 is a novel TFF1 binding protein that is important for TFF1-mediated cell migration and suggest that this interaction may be a therapeutic target in the invasion and metastasis of gastric cancer.

Keywords

TFF1 TFF1 binding protein Piezo1 Cell migration 

Abbreviations

TFF1

Trefoil factor family-1

COX

Cyclooxygenase

TXA2-R

Thromboxane A2 receptor

GC

Gastric cancer

Y2H

Yeast two-hybrid

cDNA

Complementary DNA

siRNA

Short interfering RNA

co-IP

Co-immunoprecipitation

Notes

Acknowledgments

This study was supported by National Natural Science Foundation of China (No. 81370505, 81370591 & 91229201), Key Projects for Technology Plan of Xiamen (No. 3502Z20100002) and Ministry of Health Foundation for State Key Clinical Department in China.

Conflict of interest

None.

Supplementary material

10620_2014_3044_MOESM1_ESM.tif (1.3 mb)
Supplementary Figure. A Two siRNA target sites for Piezo1 were designed and subcloned into pU6 and transfected into 293T cells to screen the silencing efficacy by western blotting. B Western blots depicting the expression of Piezo1 and TFF1 in SGC7901 and BGC-823 of cells overexpressing TFF1 (pU6-F1), TFF1 with a knockdown of Piezo1 (siP-F1), control cells transfected with an empty vector (pU6-HA) and Piezo1-knockdown cells (siP-HA). GAPHD is shown as a loading control. (TIFF 1358 kb)

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Xiao-Ning Yang
    • 1
  • Ya-Pi Lu
    • 1
  • Jing-Jing Liu
    • 1
  • Jian-Kun Huang
    • 1
  • Yun-Peng Liu
    • 1
  • Chuan-Xing Xiao
    • 1
  • Amarsanaa Jazag
    • 2
  • Jian-Lin Ren
    • 1
    Email author
  • Bayasi Guleng
    • 1
    Email author
  1. 1.Department of GastroenterologyZhongshan Hospital Affiliated to Xiamen UniversityXiamenChina
  2. 2.National Institute of Medical ResearchUlaanbaatarMongolia

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