前列腺素EP1受体通过上调粘着斑激酶磷酸化促进肝癌细胞迁移

前列腺素EP1受体通过上调粘着斑激酶磷酸化促进肝癌细胞迁移
白小明，冷静
基金项目：国家自然科学基金(81101496, 81172003);高等学校博士学科点专项科研基金新教师类资助课题基金(20113234120009) 作者简介：白小明(1979),女,讲师,主要研究方向:PGE2促进肿瘤细胞迁移与转移的机制 通信联系人：冷静(1954),男,教授,主要研究方向:PGE2促进肿瘤细胞生长与扩散的机制.

（南京医科大学病理学系,南京 210029）
摘要：目的：阐明前列腺素E2(prostaglandin E2, PGE2)通过EP1受体活化肝癌细胞Huh-7中粘着斑激酶进而促进细胞粘附与迁移的机制。方法：用PGE2、EP1受体激动剂（17-phenyltrinor Prostaglandin E2，17-PT-PGE2）、EP1受体抑制剂SC-19220等处理Huh-7细胞，通过Western blot方法检测粘着斑激酶磷酸化水平。用WST检验细胞粘附水平，用transwell小室检验细胞迁移水平。结果：17-PT-PGE2 (3 μM) 处理Huh-7细胞使粘着斑激酶FAK Y397磷酸化水平增高2倍，并使细胞粘附率与迁移率分别增高60%与40%。HEK293细胞中过表达EP1受体使FAK磷酸化水平明显增高，而EP1受体抑制剂sc-19220阻断PGE2介导的FAK磷酸化。结论：PGE2可通过EP1受体上调Huh-7细胞中FAK磷酸化水平，进而促进细胞粘附与迁移。 
关键词：病理学；EP1受体；粘着斑激酶；细胞粘附；迁移；肝癌
中图分类号：R735.7
Prostaglandin E2 EP1 Receptor Promotes Hepatocellular Carcinoma Cells Migration via regulating FAK 20 Phosphorylation
BAI Xiaoming, LENG Jing
(Department of Pathology, Nanjing Medical University, Nanjing 210029)
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 25 receptor-mediated cell adhesion and migration. We previously showed that PGE2 promotes cell adhesion and migration by activating focal adhesion kinase (FAK). This current study was designed to clarify the association between the EP1 receptor and FAK activation in HCC cells and to investigate the related signal pathway. The effects of PGE2, EP1 agonist 17-phenyl trinor-PGE2 (17-PT-PGE2) and EP1 antagonist inhibitor (sc-19220) on FAK activation were investigated by 30 treatment of Huh-7 cells. Phosphorylation of FAK Y397 was investigated by Western blotting. Cell adhesion and migration were analyzed by WST and transwell assays, respectively. 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. Meanwhile, 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. Our study suggests that the PGE2 EP1 receptor regulates FAK phosphorylation, which may regulate adhesion and migration in HCC.
Key words: Pathology; EP1 receptor; Focal Adhesion Kinase; Cell Adhesion; Migration; Hepatocellular Carcinoma