蛋白激酶C激活对口腔鳞癌细胞上皮-间质转化的影响  

The effects of PKC activation on the epithelial-mesenchymal transition of oral squamous cell cancer cells

作　　者：陈钟 郑晓丹 徐勇 

机构地区：[1]厦门医学院口腔医学系,361008

出　　处：《实用口腔医学杂志》2017年第5期674-678,共5页Journal of Practical Stomatology

基　　金：厦门医学院自然科学基金(编号:Z2013-02)

摘　　要：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-P120^(ctn)转染HN12细胞,使HN12过表达P120-连环蛋白(P120-catenin,P120^(ctn)),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、P120^(ctn)、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P120^(ctn)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。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-P120^ctn was used to transfect HN12 cells to make P120-catenin（ 120^ctn） 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, P120^ctn, 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 P120^ctn and E-cad were decreased, the cell morphology changed, mRNA 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 P120^ctn/E-cad expression and cell adhesion.

关 键 词：P120-连环蛋白(P120^ctn) 蛋白激酶C(PKC) 上皮-间质转化(EMT) 口腔鳞状细胞癌(OSCC) 

分 类 号：R739.8[医药卫生—肿瘤]