槲皮素靶向上调TLE4阻断Wnt/β-catenin信号通路抑制宫颈癌细胞的迁移及侵袭

宫颈癌是一种严重威胁女性健康的恶性肿瘤。中药有效成分槲皮素是一种潜在的宫颈癌抗肿瘤药物，然其作用机制复杂，需要进一步深入探索。结合GEO数据库及RNA-seq分析，发现Wnt通路转录抑制关键因子之一TLE4在宫颈癌中低表达，槲皮素处理后显著上调。前期实验显示：①槲皮素及过表达TLE4抑制宫颈癌细胞的迁移；②槲皮素靶向上调TLE4表达，可以阻断Wnt/β-catenin信号通路；③AR转录激活TLE4表达。因此提出科学假说：槲皮素处理宫颈癌细胞后，可能与AR蛋白分子对接，AR表达上升，转录激活TLE4表达，TLE4在核内形成转录抑制复合物，抑制Wnt/β-catenin信号通路，从而抑制宫颈癌细胞的迁移及侵袭。本项目拟从临床标本、体外细胞及体内动物实验三个层面，利用基因表达调控、Transwell、Co-IP、质谱测序、荧光共聚焦等技术阐明以上科学假说，为槲皮素治疗宫颈癌提供理论及实验依据。

Cervical cancer is a malignant tumor that seriously threatens women's health. Chinese native medicine effective component quercetin is a potential antitumor drug for cervical cancer, however, its mechanism is complicated and needs to be further explored. Combined with the GEO database and RNA-seq analysis, we found that TLE4, one of the key transcription repressors of the Wnt pathway, was low expressed in cervical cancer and significantly up-regulated after quercetin treatment. Previous experiments showed that: ①quercetin and overexpression of TLE4 inhibited the migration of cervical cancer cells; ②quercetin up-regulated the expression of TLE4, and block the Wnt/β-catenin signaling pathway; ③AR transcription activated TLE4 expression. Therefore, the scientific hypothesis was proposed: after treating cervical cancer cells with quercetin, quercetin might molecule docking AR protein, the expression of AR increased, transcription activated the expression of TLE4, TLE4 in the nucleus to form the transcription repressor complex, and inhibit the Wnt/β-catenin signaling pathway, thereby inhibiting cell migration and invasion of cervical cancer cells. This project intends to clarify the above scientific hypotheses from the three levels of clinical specimens, in vitro cells and in vivo animal experiments by using gene expression regulation, Transwell, Co-IP, mass spectrometry sequencing, fluorescence confocal, and other methods, to provide the theoretical and experimental basis for the treatment of cervical cancer with quercetin.