三叶因子3在甲状腺乳头状癌TPC-1细胞上皮间质转化中的作用及机制

Effect and mechanism of trefoil factor 3 on epithelial-mesenchymal transition of papillary thyroid carcinoma TPC-1 cells

目的探讨三叶因子3(TFF3)基因沉默对人甲状腺乳头状癌TPC-1细胞迁移、侵袭、克隆形成能力以及对上皮间质转化(EMT)的影响及机制。方法免疫组织化学技术检测甲状腺乳头状癌组织芯片中Snail与TFF3的表达。划痕实验、侵袭实验和克隆形成实验分别检测TFF3基因对TPC-1细胞迁移、侵袭和克隆能力的影响;实时定量聚合酶链反应(Real-time PCR)、免疫印迹法和免疫细胞化学染色检测上皮间质转化标志物及丝裂原活化蛋白激酶(MAPK)通路相关蛋白的变化。结果 31例甲状腺乳头状癌组织中,Snail蛋白于癌细胞胞质表达,31例癌旁组织阴性表达;Snail蛋白与TFF3表达呈正相关(r=0.8450,P<0.05)。TFF3基因沉默后,人TPC-1细胞的细胞迁移、侵袭和克隆形成能力均明显降低(P<0.01);TPC-1细胞上皮间质转化标志物及MAPK通路相关蛋白细胞外调节蛋白激酶(ERK)1/2和p-ERK1/2也显著降低(P<0.05)。结论沉默TFF3可能通过MAPK通路相关蛋白ERK1/2抑制上皮间质转化,降低TPC-1细胞迁移、侵袭和增殖能力。

Objective To explore the roles and mechanisms of silence trefoil factor 3（ TFF3） on migration,invasion,clony formation and epithelial-mesenchymal transition of thyroid papillary carcinoma TPC-1 cells. Methods Expressions of Snail and TFF3 in tissue microarrays of thyroid papillary carcinoma were analysed by immunohistochemistry.Invasion assays,wound healing assays and colony formation tests were performed to detect the effects of TFF3 on migration,invasion and clony formation of TPC-1 cells. Epithelial-mesenchymal transition（ EMT） markers and regulatory molecules were detected by Real-time PCR assay,Western blotting and immunocytochemistry in TPC-1 cells and mitogen-activated protein kinase（ MAPK） signalling pathway related proteins were examined by Western blotting and immunocytochemistry.Results In 31 cases of papillary thyroid carcinoma,Snail and TFF3 positive signals were observed in the cytoplasm of papillary throid carcinoma（ PTC）,and were negative in the para-carcinoma. The expression of Snail in papillary thyroid carcinoma showed positive correlation with the expression of TFF3 protein（ r = 0. 8450,P 0. 05）. Siliencing TFF3 expression markedly inhibited the ability of migration,invasion and clony formation in vitro via suppressing of the MAPK signalling pathway and inhibited the process of EMT（ P 0. 05 or P 0. 01）. Conclusion Knockdown of TFF3 inhibits TPC-1 cells migration,invasion clony formation and EMT related proteins epression through suppressing MAPK/ERK signalling pathway.