Hedgehog信号通路成员Sufu通过招募组蛋白去乙酰化酶3对宫颈癌上皮-间充质转化的功能影响及表观遗传机制的研究

Epithelial-mesenchymal transition (EMT) is an important factor in the development of cervical cancer. Sufu, a member of Hedgehog(Hh)-Gli pathway, can promote EMT in cervical cancer. However, the exact mechanism is still unknown. Summarizing previous studies and our work, we have hypothesized that activation of the Hh pathway enhances the binding of Sufu to histone deacetylase 3 (HDAC3) and recruits HDAC3 to regulate the epigenetic modification of the Gli transcription factor target gene E-cadherin promoter region, inhibits its expression, increases EMT, and promotes invasion and metastasis of cervical cancer. Based on the previous research, the dynamic changes of Sufu-HDAC3 binding in the nucleus will be detected by immunoprecipitation (CO-IP). Using chromatin immunoprecipitation (ChIP) and luciferase reporter gene assay to detect Sufu-HDAC3 complex in regulation of E-cadherin transcriptional activity. Finally, the cervical cancer cell line stably knocked down Sufu will be injected into the tail vein to observe the metastasis of cancer cells in vivo. The above results will help us elucidate the molecular biology mechanism of Hh-Sufu-Gli pathway in the development of cervical cancer EMT, and provide basic experimental basis for enriching and finding targeted therapy for cervical cancer.

上皮-间充质转化（EMT）是导致宫颈癌发生发展的重要因素。Hedgehog-Gli通路成员Sufu能够促进宫颈癌EMT，然而具体机制尚不清楚。总结前人和我们的研究，得出假设：Hh通路的激活增强Sufu与组蛋白去乙酰化酶3（HDAC3）的结合，招募HDAC3调控Gli转录因子靶基因E-cadherin启动子区表观遗传修饰，抑制E-cad表达、提高EMT并促进宫颈癌侵袭和转移。拟在此基础上，利用免疫沉淀（CO-IP）检测核内Sufu-HDAC3结合的动态变化；染色质免疫共沉淀（ChIP）和荧光素酶报告基因检测Sufu-HDAC3复合物对E-cadherin的转录调控；最后在整体动物水平上，尾静脉注射稳定敲低Sufu的宫颈癌细胞株至裸鼠体内，观察癌细胞的转移。上述研究结果将有助于我们阐明Hh-Sufu-Gli通路对宫颈癌EMT产生的分子生物学机制，为充实和完善靶向治疗宫颈癌提供基础实验依据。

亟需开发一种新的治疗方法来提高目前宫颈腺癌患者的预后。我们的前期研究发现，通过 HDACi可以促进PD-L1启动子区组蛋白乙酰化水平、提高染色体开放程度和PD-L1表达；说明将HDACi与PD1/PD-L1抗体联合应用，能达到抑制宫颈腺癌的作用。我们利用宫颈腺癌细胞实验进行相关分子机制研究，发现不同HDAC能够参与PD-L1转录水平的调控；利用三种HDACi处理HeLa细胞进行RNA-seq，发现不同的HDACi所调控的基因集合不同，说明HDACi所作用的基因亚群是有区别的。通过我们的研究，能够并加强阻断PD-1/PD-L1免疫抑制轴的基础理论。