LKB1对胰腺癌细胞迁移侵袭及上皮间质转化的影响及机制研究

Effects of LKB1 on Migration,Invasion and Epithelial-Mesenchymal Transition in Pancreatic Carcinoma Cells

目的研究LKB1基因对人胰腺癌细胞迁移侵袭及上皮间质转化的影响及机制。方法将LKB1过表达质粒转染至人胰腺癌细胞ASPC-1,实验分为转染试剂组、空载体组和LKB1过表达组。Western blot检测细胞转染后LKB1蛋白的表达水平;划痕实验和Transwell实验分别检测LKB1过表达对人胰腺癌细胞ASPC-1迁移、侵袭能力的影响;Western blot检测LKB1过表达对AMPKα、p-AMPKα及上皮间质转化相关蛋白E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)表达的影响。结果与转染试剂组相比,LKB1过表达质粒转染人胰腺癌细胞ASPC-1后LKB1蛋白表达水平明显增高(P<0.01),细胞迁移、侵袭能力均显著降低(P<0.01),p-AMPKα及上皮细胞标志物E-cadherin的蛋白相对表达水平增高(P<0.01),间质表型细胞标志物N-cadherin和Vimentin蛋白相对表达水平显著下降(P<0.01)。结论 LKB1抑制人胰腺癌细胞ASPC-1的迁移、侵袭和上皮间质转化,该抑制过程可能通过激活AMPK信号通路发挥作用。

Objective To investigate the effects and mechanism of LKB1 on migration, invasion and epithelial-mesenchyma] transition in human pancreatic carcinoma cells. Methods Human pancreatic carcinoma ASPC-1 cells were transfected with pcDNA3.1-LKB1. The experiment was divided into Mock, Vector and Vector-LKB1 group. The protein expression level of LKB1 was validated by Western blot. Cell migration and invasion ability was determined by the scratch tests and Transwell assay. The protein expression level of AMPKα, p-AMPKα and epithelial-mesenchymal transition（EMT）-related markers（ E-cadherin, N-cadherin and Vimentin） were assayed by Western blot analysis. Results The protein expression level of LKB1 was significantly increased in Vector-LKB1 group compared with the Mock group（P 〈 0.01 ）, and the ability of cell migration and invasion was strongly reduced（ P 〈 0.01 ）. Increased expression of p-AMPKα, epithelial marker（E-cadherin） and decreased expression of mesenchyma] markers（ N-cadherin, Vimentin） were observed in Vector-LKB1 group compared with the Mock group（ P 〈 0.01 ）. Conclusion LKB1 inhibit migration, invasion ability and epithelial-mesenchymal transition of human pancreatic carcinoma ASPC-1 cells, which may act through activating AMPK signaling pathway.