Flotillin-1激活EGFR/PI3K/Akt和NF-κB信号通路参与人肺腺癌发生与演进的研究

Our previous proteomics study of human lung adenocarcinoma suggested that up-regulation of Flotillin-1 was significantly correlated with the carcinogenesis and progression of lung adenocarcinoma, and CAP and Aurora-B were the interacting proteins of Flotillin-1. In order to eluciadate the roles and mechanisms of Flotillin-1 in the pathogenesis of human lung adenocarcinoma, in this project we will firstly establish normal human bronchial cell line and lung adenocarcinoma cell lines with Flotillin-1 expressional changes, analyze the effects of Flotillin-1 expressional changes on in vitro and in vivo cell phenotypes and biological behaviors; explore whether Flotillin-1 up-regulation activate EGFR/PI3K/Akt siganling pathway by interaction of Flotillin-1 with CAP, as well as whether Flotillin-1 up-regulation activate NF-kappaB signaling pathway through inceasing Aurora-B activity; and finally investigate wherhter Flotillin-1 up-regulation is involved in the carcinogenesis and progress of lung adenocarcinoma thruough activating the EGFR/PI3K/Akt and NF-kappaB siganling pathways.The expected results not only are helpful to elucidate the roles and molecular mechanisms of Flotillin-1 in the development and progression of human lung adenocarcinoma, but also can provide candidate molecular targets for targeted therapy of human lung adenocarcinoma.It has important theoretical significance and practical application value.

我们前期的人肺腺癌蛋白质组学研究发现，Flotillin-1表达上调与肺腺癌癌变进程密切相关，CAP和Aurora-B是Flotillin-1的相互作用蛋白。为揭示Flotillin-1在肺腺癌发病中的作用及其机制，本项目先建立Flotillin-1表达改变的正常人支气管上皮细胞系以及人肺腺癌细胞系，分析Flotillin-1表达改变对正常人支气管上皮细胞和人肺腺癌细胞表型的影响；再探讨Flotillin-1表达上调是否通过与CAP相互作用来激活EGFR/PI3K/Akt信号传导通路，以及是否通过增加Aurora-B活性来激活NF-κB信号传导通路；最后分析Flotillin-1表达上调是否通过激活这两条信号通路参与肺腺癌的发生与演进。项目研究不仅将助于揭示Flotillin-1在肺腺癌发生与演变中的作用及其分子机制，而且能为肺腺癌的靶向治疗提供分子靶标,具有重要的理论意义和实践应用价值。

目的：Flotillin-1 (Flot-1)蛋白被报道在多种肿瘤的进展中起重要作用，而flotillin-1蛋白在肺腺癌进展中的作用报道甚少。本研究旨在探讨flotillin-1蛋白在肺腺癌进展与演进中的作用。.方法：通过免疫印迹（WB）比较正常人支气管上皮细胞和5例不同肺腺癌细胞中Flotillin-1蛋白表达水平的差异。然后采用多种方法探索肺腺癌细胞系中flotillin-1的致瘤性：建立了flotillin-1过表达和flotillin-1-shRNA敲降的同源细胞系，并使用免疫印迹验证了这些同源细胞系中flotillin-1蛋白的表达差异。进一步通过细胞增殖实验、划痕愈合实验、transwell迁移实验、transwell基质胶侵袭实验，以及皮下移植瘤生长实验来评估flotillin-1蛋白在肺腺癌演进中的作用。.结果：本研究发现Flotillin-1蛋白在肺腺癌细胞中表达上调。下调flotillin-1能逆转肺腺癌在体外的恶性表型（包括增殖、迁移和侵袭），然而上调flotillin-1能促进肺腺癌在体外的恶性表型。同样地，相对空载体慢病毒感染的对照组肺腺癌细胞，flotillin-1-shRNA敲降组细胞形成了体积更小重量更轻的裸鼠皮下移植瘤。此外，肺腺癌异种移植瘤模型中，对照组肺腺癌细胞相对flotillin-1-shRNA敲降组肺腺癌细胞表现出了更明显的致瘤效应和恶性表型。抑制flotillin-1的表达水平能抑制NF-κB入核。