hsa_circ_001634/miR-194/Wnt轴参与胃癌发生发展的机制研究

Based on our preliminary data, miR-194 was identified as an oncogenic miR in gastric cancer (GC). It promoted cell growth and migration in GC cells. According to the bioinformatic analysis and preliminary results, we propose a new regulatory mechanism of miR-194: miR-194, regulated by hsa_circ_001634, contributes to the GC tumorigenesis by activating Wnt signaling pathway via targeting SUFU. We intend to elucidate the mechanism by using different transfection combination, interference, luciferase reporter system, CHIP, confocal immunofluorescence and biological function tests. (1) We would like to explore the biological function of hsa_circ_001634 and miR-194 in gastric cells and determine if hsa_circ_001634 is an upstream regulatory factor for miR-194. (2)We will characterize the hsa_circ_001634/miR-194/Wnt axis signaling pathway in GC tumorigenesis. (3) We would confirm SUFU, AXIN2 are the target genes of miR-194. The project will ultimately reveal a new regulatory mechanism of miR-194 and clarify the function of hsa_circ_001634/miR-194/Wnt axis signaling pathway in GC tumorigenesis. It will provide novel circRNA, miR molecular markers and gene targets for clinical diagnosis and treatment of GC.

预实验中我们发现miR-194在胃癌中促进细胞的生长和迁移，是一个致癌miR。根据生物信息学分析及预实验结果，提出了一个新的miR-194调控机制：即miR-194在hsa_circ_001634调控下，通过SUFU等靶基因激活Wnt信号通路促进胃癌发生和发展。本项目拟利用各种转染、荧光素酶报告系统、CHIP、共聚焦免疫荧光、体内外生物学功能等实验验证这一机制。最终探明（1）hsa_circ_001634和miR-194 在胃细胞中的生物学功能，hsa_circ_001634是miR-194的上游调控因子。（2）hsa_circ_001634/miR-194/Wnt轴信号通路的作用机制。（3）SUFU、AXIN2等是miR-194的靶基因。旨在阐明hsa_circ_001634/miR-194/Wnt轴信号通路在胃癌演进中的作用机制，为临床胃癌诊治提供新的分子标志物和基因靶点。

本项目探讨了hsa_circ_001634/miRNA-194/Wnt轴信号通路在胃癌发生及转移中的作用机制，结果显示 miRNA-194 在胃癌组织及细胞中表达明显增高,提示 miRNA-194 具有一定的促进胃癌发生和发展的作用；转染 miRNA-194 抑制剂 后，胃癌细胞株 BGC823 的增殖、侵袭等生物学功能分别被抑制；体内成瘤实验证实， miRNA-194 抑制剂具有一定的抑制BGC823细胞肿瘤形成能力；通过生物信息学、基因表达谱芯片及双荧光素酶活性检测以及免疫印迹实验证实 SUFU 是 miRNA-194 的直接靶基因；应用miRNA-194 抑制剂，Wnt通路活性被抑制;体外实验证实SUFU 能有效逆转 miRNA-194 inh 对 BGC823 细胞的增殖、侵袭、迁移及Wnt通路抑制作用; hsa_circ_001634在胃癌中通过sponge miRNA-194,抑制Wnt通路活性。由此，本项目揭示了 miR-194在hsa_circ_001634调控下，通过SUFU等靶基因激活Wnt信号通路促进胃癌发生和发展的机制。此项目的开展不仅有助于阐明胃癌发生发展的新分子机制，还为临床抗胃癌转移提供新的靶点和治疗策略。