Prostaglandin E2 receptor EP1-mediated phosphorylation of focal adhesion kinase enhances cell adhesion and migration in hepatocellular carcinoma cells

Bai,Xiaoming; Wang,Jie; Zhang,Li; Ma,Juan; Zhang,Hai; Xia,Shukai; Zhang,Min; Ma,Xiuping; Guo,Yan; Rong,Rong; Cheng,Shanyu; Shu,Wei; Wang,Yipin; Leng,Jing

doi:10.3892/ijo.2013.1859

Prostaglandin E2 receptor EP1-mediated phosphorylation of focal adhesion kinase enhances cell adhesion and migration in hepatocellular carcinoma cells

Authors:
- Xiaoming Bai
- Jie Wang
- Li Zhang
- Juan Ma
- Hai Zhang
- Shukai Xia
- Min Zhang
- Xiuping Ma
- Yan Guo
- Rong Rong
- Shanyu Cheng
- Wei Shu
- Yipin Wang
- Jing Leng
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Affiliations: Cancer Center, Department of Pathology, Nanjing Medical University, Nanjing 210029, P.R. China, Department of Neurology, The Affiliated Hospital of Xuzhou Medical College, Xuzhou 221000, P.R. China, Department of Pathology, The First Affiliated Hospital with Nanjing Medical University, Nanjing 210029, P.R. China, Department of Periodontal, Institute of Stomatology, The Stomatological Hospital Affiliated to Nanjing Medical University, Nanjing 210029, P.R. China
Published online on: March 20, 2013 https://doi.org/10.3892/ijo.2013.1859
Pages: 1833-1841

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Abstract

The prostaglandin E2 (PGE2) EP1 receptor has been implicated in hepatocellular carcinoma (HCC) cell invasion. However, little is known about the mechanisms of EP1 receptor-mediated cell adhesion and migration. We previously showed that PGE2 promotes cell adhesion and migration by activating focal adhesion kinase (FAK). The present study was designed to elucidate the association between the EP1 receptor and FAK activation in HCC cells and to investigate the related signaling pathways. The effects of PGE2, EP1 agonist 17-phenyl trinor-PGE2 (17-PT-PGE2), PKC and EGFR inhibitors on FAK activation were investigated by treatment of Huh-7 cells. Phosphorylation of FAK Y397 and c-Src Y416 was investigated by western blotting. Cell adhesion and migration were analyzed by WST and transwell assays, respectively. Protein kinase C (PKC) activity was measured with a PKC assay kit. The results showed that 17-PT-PGE2 (3 µM) increased FAK Y397 phosphorylation by more than 2-fold and promoted cell adhesion and migration in Huh-7 cells. In transfected 293 cells, expression of the EP1 receptor was confirmed to upregulate FAK phosphorylation, while the EP1 receptor antagonist sc-19220 decreased PGE2-mediated FAK activation. PKC activity and c-Src Y416 phosphorylation were enhanced after 17-PT-PGE2 treatment. Both PKC and c-Src inhibitor suppressed the 17-PT-PGE2-upregulated FAK phosphorylation, as well as 17-PT-PGE2-induced cell adhesion and migration. In addition, exogenous epidermal growth factor (EGF) treatment increased FAK phosphorylation. The EGF receptor (EGFR) inhibitor also suppressed 17-PT-PGE2-upregulated FAK phosphorylation. Our study suggests that the PGE2 EP1 receptor regulates FAK phosphorylation by activating the PKC/c-Src and EGFR signal pathways, which may coordinately regulate adhesion and migration in HCC.

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