ABSTRACT

FGF13, a member of the FGF subfamily, has been found to be highly expressed in cancer cells such as prostate cancer, melanoma, glioma and multiple myeloma. However, the mechanism of FGF13 function during cancer cell proliferation remains to be unexplored, especially Non-small cell lung cancer (NSCLC). In this study, the cell proliferation effect of FGF13 on A549 cells was checked by CCK-8, clone formation, Ki67 immunofluorescence staining and Flow Cytometry assay. Localization of FGF13 within A549 cells was performed with confocal laser scanning microscope. The protein variations and interaction were measured by western blotting and co-immunoprecipitation analysis. It showed that FGF13 was mainly distributed in the cytoplasm and exhibited a high expression level in A549 cells. High expression of FGF13 activated AKT-GSK3 signaling pathway, and inhibited the activity of p21 and p27. Thus, FGF13 enhanced the process of transition from G1 to S phase and promoted A549 cells proliferation. Furthermore, the interaction between FGF13 and SHCBP1 was confirmed. Meanwhile, FGF13 and SHCBP1 had a cooperative effect to accelerate the cell cycle progression, especially the ability to promote cell proliferation is significantly enhanced via protein interaction. Hence, we conclude that FGF13 played a positive regulation role during A549 cells proliferation. FGF13 interacted with SHCBP1 to facilitate cell cycle progression, providing new insights into deep understanding of non-small cell lung cancer mechanisms of proliferation and regulation function of FGF13.

摘要

FGF13是FGF亚家族的成员，已被发现在前列腺癌、黑色素瘤、神经胶质瘤和多发性骨髓瘤等癌细胞中高表达。然而，FGF13在癌细胞增殖过程中的作用机制仍有待探索，尤其是非小细胞肺癌（NSCLC）。本研究通过CCK-8、克隆形成、Ki67免疫荧光染色和流式细胞术检测FGF13对A549细胞的细胞增殖作用。使用共焦激光扫描显微镜对 A549 细胞内的 FGF13 进行定位。通过蛋白质印迹和免疫共沉淀分析来测量蛋白质变异和相互作用。表明FGF13主要分布于细胞质,在A549细胞中呈现高表达水平。FGF13的高表达激活AKT-GSK3信号通路，并抑制p21和p27的活性。因此，FGF13增强了G1期向S期转变的过程并促进A549细胞增殖。此外，还证实了FGF13和SHCBP1之间的相互作用。同时，FGF13和SHCBP1协同作用，加速细胞周期进程，特别是通过蛋白质相互作用，促进细胞增殖的能力显着增强。因此，我们得出结论，FGF13在A549细胞增殖过程中发挥了正向调节作用。FGF13与SHCBP1相互作用促进细胞周期进展，为深入理解非小细胞肺癌增殖机制和FGF13的调节功能提供了新的见解。