蛋白激酶C激活对口腔鳞癌细胞上皮-间质转化的影响

发布时间：2018-10-05 17:19

【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(P0.05)。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。
[Abstract]:Aim: to explore the effect of activated protein kinase (C (protein kinase) on epithelial-interstitial transformation (epithelial-mesenchymal transition,EMT) of oral squamous carcinoma cells (OSCC) by using tongue cancer cell line HN12. Methods: HN12 cells were transfected with plasmid pCMV6-AC-GFP-P120~ (ctn), and HN12 was overexpressed by P120-cateninine (P120-cateninine) P120 ~ (ctn), and then added into phorbol 12-myristate 13-acetate PMA. The mRNA and protein expression of PKC,P120~ (ctn) E-cad before and after PMA treatment were detected by real-time fluorescence quantitative PCR,Western blot. The ability of cell migration and invasion was detected by Transwell cell invasion and cell migration assay. Results: when PKC was activated by PMA, the cell morphology was changed, and the expression of E-cadherin (E-cadherin E-cad) was decreased, and the mRNA and protein expression of N-cadherin (N-cadherin N-cad) and vimentin (Vimentin,Vim) were increased significantly. The ability of migration and invasion of tumor cells was significantly increased (P0.05). ConclusionPKC may play a role in the migration and invasion of oral squamous carcinoma cells by regulating the expression of P120 ~ (ctn) through phosphorylation, which may play an important role in the regulation of cell adhesion and promote the migration and invasion of oral squamous carcinoma cells.
【作者单位】：厦门医学院口腔医学系;
【基金】：厦门医学院自然科学基金(编号:Z2013-02)
【分类号】：R739.8

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