ABSTRACT
Objective:To study the effects of activating protein kinase C (PKC)on oral squamous-cell cancer(OSCC) cell epithelial-mesenchymal transition(EMT).Methods:Plasmid pCMV6-AC-GFP-P120ctn was used to transfect HN12 cells to make P120-catenin (120ctn) overexpress and thereafter PKC activator PMA (phorbol 12-myristate 13-acetate) was added to the culture.Real-time fluorescent quantitative PCR and Western blot were adopted to test mRNA and protein expression of PKC,P120ctn,E-cadherin(E-cad),N-cadherin(N-cad) and Vimentin(Vim).Transwell cell invasion and cell migration assay were used to test the invasion and migration capacity before and after the activation.Results:When PKC was activated by PMA,the expression of P120ctn and E-cad were decreased,the cell morphology changed,nRNA and protein expression of mesenchymal marker protein N-cad and Vim increased significantly.Meanwhile,the migration and invasion capacity of the tumor cells increased significantly(P ＜ 0.05).Conclusion:In OSCC cells,PKC may be involved in promoting EMT and the metastasis and invasion by adjusting P120ctn/E-cad expression and cell adhesion.
ABSTRACT
Objective:To study the expression of IQ motif containing GTPase activating protein 1 (IQGAP1 )in oral squamous cell cancer(OSCC)tissue,and to explore the effects of IQGAP1 on cell proliferation and invasion as well as its underlying mechanism. Methods:Expression levels of IQGAP1 in tumor and adjacent normal tissues were examined by western blot and RT-PCR.OSCC cell line SCC-4 cells was transfected with the recombinant plasmid-pcDNA3.1 -IQGAP1 by lipofectamine,and then treated with an Akt in-hibitor.The phosphorylation of Akt,cell proliferation and invasion were detected by western blot,MTT assay and Transwell invasion as-say respectively.Results:Protein and mRNA expression levels of IQGAP1 were higher in cancer tissue than in adjacent normal tissue (P <0.05).Transfection of pcDNA3.1 -IQGAP1 increased IQGAP1 expression,enhanced the capability of cell proliferation and inva-sion (P <0.05),increased p Akt level in the cells.Preconditioning with an Akt inhibitor reduced p Akt level.Furthermore,silencing Akt pathway blocked the increase of cell proliferation and invasion induced by IQGAP1 overexpression(P <0.05).Conclusion:IQ-GAP1 overexprission can mediate the ability of proliferation and invasion of OSCC cells by regulating the activation of Akt pathway.