CKLF1通过激活PLCγ/FAK信号途径促进SH-SY5 Y细胞迁移

CKLF1 induces SH-SY5Y cell migration via PLCγ/FAK signaling pathway

目的考察趋化素样因子1(chemokine-like factor 1,CKLF1)对神经系统细胞系SH-SY5Y细胞迁移运动的影响,并进一步研究其分子机制。方法采用CELLocate细胞定位法研究CKLF1在不同时间点对SH-SY5Y细胞迁移的影响;采用蛋白免疫印迹法观察CKLF1是否能够增强与细胞迁移密切相关的局部粘着斑激酶(focal adhesion kinase,FAK)Tyr-397位点的磷酸化水平,并进一步研究抑制FAK上游分子PLCγ对这一作用的影响;采用Boyden趋化小室法观察抑制FAK或PLCγ对CKLF1趋化活性的影响。结果 CKLF1对SH-SY5Y细胞具有趋化作用,能够诱导其迁移运动,并能明显增强FAK-pY397位点的磷酸化水平,而加入PLCγ抑制剂U73122则减弱了CKLF1诱导的FAK磷酸化水平的升高。此外,FAK抑制剂PF-573228和PLCγ抑制剂U73122均可明显抑制CKLF1的趋化活性。结论趋化因子CKLF1能够通过激活PLCγ/FAK信号途径,促进SHSY5Y细胞的迁移运动。

Aim To investigate the role of chemokine-like factor 1 （ CKLF1 ） in SH-SY5 Y cell migration and its molecular regulatory mechanism. Methods SH-SY5Y cells were stimulated with CKLF1 for 0. 5 h, 2 h, 8 h and 24 h, respectively. The migration distance and the percentage of migration cells were recorded by CELLocate analysis. The phosphorylation of focal ad-hesion kinase （ FAK） at Tyr-397 site was detected by Western blot analysis. By chemotaxis assays, we con-firmed the chemotaxis of CKLF1. Furthermore, FAK inhibitor PF-573228 and PLCγ inhibitor U73122 were used for the research of molecular regulatory mecha-nisms involved. Results CKLF1 promoted cell migra-tion and induced a strong increase in the phosphoryla-tion level of FAK-pY397 , which were significantly at-tenuated by the presence of U73122 （ a specific inhibi-tor for PLCγ） . In addition, the chemotaxis of CKLF1 was obviously blocked by the FAK inhibitor PF-573228 . Conclusion CKLF1 induces SH-SY5 Y cell migration via PLCγ/FAK signaling pathway.