EGF上调TRPM7诱导肿瘤细胞迁移的分子机制研究  

Molecular Mechanisms for TRPM7 Involvement in EGF-induced Migration of Lung Adenocarcinoma Cells

作　　者：杲海霞[1] 乔晓温 敦洁宁[3] 董明纲[4] 宋桂芹[4] 徐志伟[4] 张海林[1] 

机构地区：[1]河北医科大学药理学教研室,河北石家庄050000 [2]河北省辛集市中医院儿科,河北辛集052360 [3]河北医科大学药学系药剂教研室,河北石家庄050000 [4]河北北方学院,河北张家口075000

出　　处：《河北北方学院学报（自然科学版）》2014年第2期59-63,共5页Journal of Hebei North University:Natural Science Edition

基　　金：国家自然科学基金青年基金项目(No.81201642)

摘　　要：目的探讨表皮生长因子(EGF)上调顺时受体电位M7(TRPM7)以及TRPM7参与EGF诱导肿瘤细胞迁移的分子机制。方法 Western blot技术用于检测Akt和vimentin的蛋白表达,钙成像技术用于细胞内钙离子浓度的检测,全细胞膜片钳技术用于记录TRPM7电流,激光共聚焦技术检测细胞骨架蛋白F-actin的形态。结果 Western blot的结果显示EGF显著增加A549细胞Akt的磷酸化水平;PI3K阻断剂wortmanin可显著抑制EGF上调TRPM7电流的作用;EGF孵育A549细胞可上调vimentin的蛋白表达水平,并改变细胞骨架蛋白F-actin的荧光强度以及形态;稳定转染TRPM7-shRNA可抑制高钙引起的细胞内钙升高及EGF对细胞形态及F-actin表达的作用。结论 EGF可能通过PI3K-Akt信号通路上调TRPM7膜蛋白表达和通道功能,抑制钙离子内流,从而影响EMT的产生和F-actin的聚合,最终诱导细胞迁移。Objective To study the molecular mechanisms for TRPM7 involvement in EGF-induced migration of lung adenocarcinoma cells.Methods Western blot was used to detect the protein expression of Akt and vimentin,while calcium photometry was utilized to measure the changes of intracellular Ca2+concentration and whole cell patch clamp technique was for studying TRPM7 currents.Meanwhile,con-focal microscopy was for investigating F-actin. Results Results of Western blot showed that EGF in-creased the protein expression of phosphorylated Akt (p-Akt)in A549 cells significantly,but wortmanin (PI3K inhibitor)decreased EGF-induced up-regulation of TRPM7.EMT and assembly of F-actin were al-so important for EGF-induced migration of A549 cells and regulation of TRPM7.Some morphology chan-ges appeared in EGF-treated A549 cells,which increased the protein expression of vimentin (typical pro-tein of EMT).increased the density of F-actin and induced reorganization of actin cytoskeleton from long to short actin filaments.TRPM7-shRNA inhibited the effects of EGF on reorganization of actin filaments and decreased Ca2+ influx induced by high external Ca2+ in A549 cell.Conclusion EGF might up-regulate TRPM7 through PI3K-Akt signaling pathway,which increased influx of Ca2+,EMT and assembly of F-actin,and then enhanced migration of cancer cells.

关 键 词：表皮生长因子 顺时受体电位 M7 肿瘤细胞 迁移 

分 类 号：R349.5[医药卫生—基础医学]