蛋白激酶C激活对口腔鳞癌细胞上皮-间质转化的影响

摘要

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.%目的:利用舌癌细胞系HN12探索激活蛋白激酶C(protein kinase C,PKC)对口腔鳞癌细胞上皮-间质转化(epithelial-mesenchymal transition,EMT)的影响.方法:采用质粒pCMV6-AC-GFP-P120ctn转染HN12细胞,使HN12过表达P120-连环蛋白(P120-catenin,P120ctn),再加入蛋白激酶C(protein kinase C,PKC)激活剂佛波酯(phorbol 12-myristate 13-acetate,PMA),采用实时荧光定量PCR、Western blot检测PMA处理前后PKC、P120ctn、E-cad的mRNA和蛋白表达,通过Transwell细胞侵袭及细胞迁移试验等方法检测细胞的迁移和侵袭能力.结果:当PKC被PMA活化时,细胞形态发生改变,细胞中P120ctn的表达显著降低,E-钙黏蛋白(E-cadherin,E-cad)也随之降低,间质标记蛋白N-钙黏蛋白(N-cadherin,N-cad)和波形蛋白(Vimentin,Vim)的mRNA和蛋白表达水平显著升高,肿瘤细胞的迁移和侵袭能力显著提高(P＜0.05).结论:PKC可能通过磷酸化调节P120ctn的表达,参与细胞黏附的调控,促进EMT,在口腔鳞癌细胞的迁移和侵袭过程中发挥作用.