The Functional Study Of PKCζ In The Chemotaxis Of Human Non-Small Cell Lung Cancer

Objective: In this study, the aim was to detect the function of PKCζin the chemotaxis of human non-small cell lung cancer induced by EGF or EGFR, so as to explore the anti-tumor target for the therapy of metastasis of lung cancer and clinical application.Methods: Human non-small cell lung cancer (NSCLC) cell lines A549 and H1299 were used as objects, the expression of EGFR and PKCζwas detected by Western blotting and RT-PCR; the chemotactic capacity of A549 induced by EGF was contrasted with which induced by SDF-1α/CXCL12 during chemotaxis assay; treatment with different inhibitors of PKC and myr-pseudosubstrate of PKCζ, the functions of different isotypes of PKC were observed in the chemotaxis with or without EGF (as a negative control) stimulation; PKCζtranslocation and F-actin polymerization were analysed by immunofluorescent staining technique; the expression level of EGFR was quantified by FACS before and after myr-pseudosubstrate treatment; the content of F-actin was construed through Alexa Fluor 568 methanol extraction/phalloidin staining and fluorescence analysis at 578nm excitation and 600nm emission; the number of attached cells was counted by microscopy after incubation with inhibitors or myr-pseudosubstrate followed by EGF stimulation.Results: EGF-induced chemotaxis indexes were about 3.6-fold more than those induced by CXCL12. Among different inhibitors, chelerythrine chloride, an inhibitor of all PKC isotypes, and PKCζmyr-pseudosubstrate...