食管癌相关基因2抑制细胞侵袭迁移的机制研究  

Mechanism underlying the ECRG2 inhibition of cell invasion and migration

作　　者：原发家[1] 毕炀辉 杜斌[1] 王文健[1] 聂守民 李彦红[2] 赵瑞君[1] 刘静[3] 

机构地区：[1]山西医科大学基础医学院寄生虫教研室,太原030001 [2]山西医科大学第二医院心内科 [3]山西医科大学第一临床医学院

出　　处：《中国药物与临床》2015年第3期301-304,共4页Chinese Remedies & Clinics

基　　金：国家自然科学基金(81272189)

摘　　要：目的本研究旨在明确食管癌相关基因(ECRG)2基因抑制肿瘤细胞侵袭迁移的分子机制。方法利用HT1080-Tet-On可诱导表达系统,采用基因敲除技术、免疫共沉淀、体外Matrigel transwell实验等技术,深入研究ECRG2基因抑制肿瘤细胞侵袭迁移的分子机制。结果 1敲除HT1080-ECRG2-Tet-On可诱导表达细胞株中的ECRG2后,加入多西环素诱导ECRG2蛋白高度表达,撤除多西环素后ECRG2蛋白表达降低;2HT1080-ECRG2-Tet-On-3′UTR-si可诱导细胞株中尿激酶型纤溶酶原激活剂(u PA)分解尿激酶型纤溶酶原激活剂受体(u PAR)产生分解后的u PAR(D2D3);而加入多西环素诱导ECRG2表达后,ECRG2通过与u PA/u PAR直接结合,抑制u PA对u PAR的分解作用;3HT1080-ECRG2-Tet-On-3′UTR-si可诱导细胞株中u PAR/甲酰肽样受体(FPRL1)的结合作用显著增强,而添加多西环素诱导ECRG2表达细胞中u PAR/FPRL1的结合作用被完全阻断,撤除多西环素后结合作用又恢复;4敲除HT1080-ECRG2-Tet-On-3′UTR-si可诱导细胞株中FPRL1,细胞的侵袭迁移能力降低。结论 ECRG2通过u PA/u PAR/FPRL1通路抑制肿瘤细胞侵袭迁移。Objective To determine the molecular mechanisms underlying the inhibitory action of ECRG2 a gainst tumor cell invasion and migration. Methods HT1080-Tet-On inducible expression system, gene knockout techniques, co-immunoprecipitation, and matrigel transwell in vitro were used to explore the tumor resistance of gene ECRG2. Results 1 When ECRG2 was knocked out from cell line HT1080-ECRG2-Tet-On, adding doxcyclin may result in high expression of ECRG2 protein, but the expression was reduced after removing doxcyclin. 2 u PA could decompose u PAR to generate cleaved u PAR（D2D3） in the HT1080-ECRG2- Tet-On-3′UTR-si inducible cell line;whereas the highly expressed ECRG2 induced by doxcyclin could inhibit u PA decomposing u PAR by binding to u PA/u PAR directly. 3The binding of u PAR / FPRL1 in HT1080-ECRG2- Tet-On-3′UTR-si cell lines could be significantly enhanced; and the highly expressed ECRG2 induced by doxcyclin could block the binding ability between u PAR and FPRL1, whereas the binding ability could resume by removing doxcyclin. 4 When FPRL1 was knocked out from HT1080- ECRG2- Tet-On-3＇ UTR-si-inducible cell line, a reduction in cell invasion and migration was noted. Conclusion ECRG2 could inhibit tumor cell invasion and migration through u PA / u PAR / FPRL1 pathway.

关 键 词：肿瘤浸润 癌基因 ECRG2 UPAR FPRL1 

分 类 号：R735.7[医药卫生—肿瘤]