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

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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.

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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

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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.

Author information

Authors and Affiliations

  1. Department of Gastroenterology, Zhongshan Hospital Affiliated to Xiamen University, 201 Hubin South Road, Xiamen, 361004, Fujian Province, China

    Xiao-Ning Yang, Ya-Pi Lu, Jing-Jing Liu, Jian-Kun Huang, Yun-Peng Liu, Chuan-Xing Xiao, Jian-Lin Ren & Bayasi Guleng

  2. National Institute of Medical Research, Tsend-Ayush Street, 3rd General Hospital, Ulaanbaatar, Mongolia

    Amarsanaa Jazag

Authors
  1. Xiao-Ning Yang
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  2. Ya-Pi Lu
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  5. Yun-Peng Liu
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  8. Jian-Lin Ren
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  9. Bayasi Guleng
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Corresponding authors

Correspondence to Jian-Lin Ren or Bayasi Guleng.

Additional information

Xiao-Ning Yang and Ya-Pi Lu contributed equally to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

10620_2014_3044_MOESM1_ESM.tif

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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Yang, XN., Lu, YP., Liu, JJ. et al. Piezo1 Is as a Novel Trefoil Factor Family 1 Binding Protein that Promotes Gastric Cancer Cell Mobility In Vitro. Dig Dis Sci 59, 1428–1435 (2014). https://doi.org/10.1007/s10620-014-3044-3

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  • DOI: https://doi.org/10.1007/s10620-014-3044-3

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