PI3K/Akt与MEK/ERK信号通路间的“串话”对肺癌细胞自噬调控的机制研究

The research of crosstalk between different signaling pathways in malignant cells becomes more concern. The regulation of autophagy by crosstalk between PI3K/Akt and MEK / ERK signaling pathways in malignant cells has been rarely reported and has not been reported in lung cancer cells. In our previous study, we found that the crosstalk between PI3K/Akt and MEK / ERK signaling pathway in lung cancer cells, and it is closely related with cell types, gene mutation and other factors. This crosstalk affected autophagy in lung cancer cells by cross-activation or cross-inhibition. So we assumed that the mechanism of crosstalk is an important factor of autophagy regulation in lung cancer cells. Based on this, the study intends to use cancer tissue, cell culture in vitro as well as xenografts in nude mice by flow cytometry, qRT-PCR, Western blot, a small animal in vivo imaging methods, histology, to explore the molecular mechanism of the "cross-talk" between PI3K/Akt and MEK / ERK signaling pathway in different levels.Moreover, It will reveal potential mechanisms accounting for crosstalk-induced autophagy. Our results will also deepen the understanding of the mechanisms of lung cancer and provide a strong scientific rationale for effective individualized treatment of lung cancers.

恶性肿瘤信号通路间的"串话"研究正在被引起关注，有关PI3K/Akt与MEK/ERK信号通路的"串话"对恶性肿瘤细胞自噬调控的研究少有报告，而与肺癌细胞自噬的调控机制尚未见报告。我们前期试验发现肺癌细胞中存在PI3K/Akt与MEK/ERK信号通路间的"串话"现象，并且与细胞类型、基因突变等因素密切相关，并影响着自噬对肺癌细胞的双向调节作用。由此推测不同的"串话"机制对肺癌细胞自噬的调控是一个重要因素。基于此，本研究拟采用临床病理、体外癌细胞培养以及裸鼠肺癌皮下移植瘤模型等手段，通过流式细胞术、qRT-PCR、Western blot、小动物活体成像等方法，从组织学、细胞学、分子生物学层面，探讨PI3K/Akt与MEK/ERK信号通路间"串话"的分子机制，揭示其对肺癌细胞自噬的调控规律，加深对肺癌发生机制的理解，为个体化治疗提供新的思路和理论依据。

恶性肿瘤信号通路间的“串话”研究正在被引起关注，有关PI3K/Akt与MEK/ERK信号通路的“串话”对恶性肿瘤细胞自噬调控的研究少有报告，而与肺癌细胞自噬的调控机制尚未见报告。本研究采用了临床病理、体外癌细胞培养以及裸鼠肺癌皮下移植瘤模型等手段，通过流式细胞术、qRT-PCR、Western blot、小动物活体成像等方法，从组织学、细胞学、分子生物学层面，探讨了PI3K/Akt与MEK/ERK信号通路间“串话”的分子机制。. 我们检测到了PI3K抑制剂BKM120能够，上调自噬的标志物LC-3蛋白的表达，特别是在对BKM120敏感的细胞株中更明显。当其与溶酶体蛋白酶抑制剂氯喹联用时可进一步增强型LC3蛋白的表达，提示自噬对BKM120抑制细胞生长具有保护作用。BKM120增加p-ERK的表达。当抑制或基因沉默ERK时，BKM120上调LC3的作用被减弱。这表明MEK / ERK的活化有助于BKM120诱导authophagy。在小鼠异种移植模型中，我们发现，PI3K抑制剂BKM120联合MEK抑制剂PD0325901协同抑制肺癌细胞的生长。因此，本研究不仅揭示了机制占BKM120诱导自噬，并提议通过联合使用溶酶体蛋白酶抑制剂或MEK抑制剂阻断自噬从而加强BKM120治疗非小细胞肺癌疗效的方法。 . 研究揭示PI3K/Akt与MEK/ERK信号通路的“串话”对肺癌细胞自噬调控的机制，加深对肺癌发生机制的理解，为个体化治疗提供新的思路和理论依据。