Abstract：
BACKGROUND: Gastric cancer (GC) is a leading cause of cancer-associated death worldwide. Its molecular mechanisms, especially concerning autophagy and various signaling pathways, are not fully understood. Fatty Acid Binding Protein 6 (FABP6) and RE1 Silencing Transcription Factor (REST) emerge as potential key players in this context. This study sought to analyze the functional relationship of FABP6 and REST concerning autophagy and their implications on the Akt/mTOR signaling pathway within GC cells.
METHODS: A comprehensive bioinformatics approach was used to identify key prognostic markers in GC. The effects of FABP6 and REST on autophagy along with Akt/mTOR signaling pathways were analyzed by techniques including Western blotting (WB), flow cytometry, Transwell assay, dual luciferase reporter assay, and others.
RESULTS: FABP6 was identified as overexpressed in GC, linked with poor prognosis. FABP6 silencing reduces GC cell proliferation, induces S- and G2-phase arrest, and downregulates cyclins CDK2 and CDK4. It also inhibited GC cell invasion/migration and autophagy, effects that were counteracted by MG132. When combined with PI3K inhibitor LY294002c, FABP6 knockdown showed synergistic anti-proliferative effects, modulating the Akt/mTOR pathway. Besides, the transcription factor REST has been shown to directly regulate FABP6 expression, affecting autophagy and the Akt/mTOR signaling pathway in a FABP6-dependent manner.
CONCLUSIONS: REST positively regulates autophagy and negatively affects the Akt/mTOR signaling pathway in GC cells in a FABP6-dependent manner, providing valuable insights into regulatory networks involving FABP6 and REST.
METHODS: A comprehensive bioinformatics approach was used to identify key prognostic markers in GC. The effects of FABP6 and REST on autophagy along with Akt/mTOR signaling pathways were analyzed by techniques including Western blotting (WB), flow cytometry, Transwell assay, dual luciferase reporter assay, and others.
RESULTS: FABP6 was identified as overexpressed in GC, linked with poor prognosis. FABP6 silencing reduces GC cell proliferation, induces S- and G2-phase arrest, and downregulates cyclins CDK2 and CDK4. It also inhibited GC cell invasion/migration and autophagy, effects that were counteracted by MG132. When combined with PI3K inhibitor LY294002c, FABP6 knockdown showed synergistic anti-proliferative effects, modulating the Akt/mTOR pathway. Besides, the transcription factor REST has been shown to directly regulate FABP6 expression, affecting autophagy and the Akt/mTOR signaling pathway in a FABP6-dependent manner.
CONCLUSIONS: REST positively regulates autophagy and negatively affects the Akt/mTOR signaling pathway in GC cells in a FABP6-dependent manner, providing valuable insights into regulatory networks involving FABP6 and REST.
摘要：
背景：胃癌（GC）是全球癌症相关死亡的主要原因。其分子机制，特别是关于自噬和各种信号通路，没有完全理解。脂肪酸结合蛋白6（FABP6）和RE1沉默转录因子（REST）在这种情况下成为潜在的关键参与者。本研究旨在分析FABP6和REST与自噬的功能关系及其对GC细胞内Akt/mTOR信号通路的影响。
方法：使用综合生物信息学方法来鉴定GC的关键预后标志物。通过包括蛋白质印迹（WB）在内的技术分析了FABP6和REST对自噬以及Akt/mTOR信号通路的影响。流式细胞术,Transwell分析，双荧光素酶报告分析，和其他人。
结果：FABP6在GC中被鉴定为过表达，与预后不良有关。FABP6沉默降低GC细胞增殖，引起S-和G2-阶段停滞，并下调细胞周期蛋白CDK2和CDK4。它还抑制GC细胞侵袭/迁移和自噬，MG132抵消的影响。当与PI3K抑制剂LY294002c联合使用时，FABP6敲低显示协同抗增殖作用，调节Akt/mTOR途径。此外,转录因子REST已被证明直接调节FABP6的表达，以FABP6依赖性方式影响自噬和Akt/mTOR信号通路。
结论：REST正向调节自噬，并以FABP6依赖性方式对GC细胞中的Akt/mTOR信号通路产生负面影响，为涉及FABP6和REST的监管网络提供有价值的见解。
方法：使用综合生物信息学方法来鉴定GC的关键预后标志物。通过包括蛋白质印迹（WB）在内的技术分析了FABP6和REST对自噬以及Akt/mTOR信号通路的影响。流式细胞术,Transwell分析，双荧光素酶报告分析，和其他人。
结果：FABP6在GC中被鉴定为过表达，与预后不良有关。FABP6沉默降低GC细胞增殖，引起S-和G2-阶段停滞，并下调细胞周期蛋白CDK2和CDK4。它还抑制GC细胞侵袭/迁移和自噬，MG132抵消的影响。当与PI3K抑制剂LY294002c联合使用时，FABP6敲低显示协同抗增殖作用，调节Akt/mTOR途径。此外,转录因子REST已被证明直接调节FABP6的表达，以FABP6依赖性方式影响自噬和Akt/mTOR信号通路。
结论：REST正向调节自噬，并以FABP6依赖性方式对GC细胞中的Akt/mTOR信号通路产生负面影响，为涉及FABP6和REST的监管网络提供有价值的见解。
