circNEK6上调HNF4A/NEK6/Cyclin D2表达的分子机制及其在促进贲门腺癌发生发展中的作用

In our previous study, it was found that circNEK6 was up-regulated in gastric cardiac adenocarcinoma (GCA) cancerous tissues compared with their paired normal tissues. Depletion of CircNEK6 led to the inhibition of HNF4A, NEK6 and Cyclin D2, and the up-regulation of miR-532-5p、miR-339-5p、miR-665, resulting in decreased potential of colony formation of GCA cells. Especially, NEK6 is the host gene of circNEK6, HNF4A, NEK6 and Cyclin D2 are targets of miR-532-5p、miR-339-5p、miR-665, and HNF4A are the potential transcription factor of NEK6 and Cyclin D2 gene. Hence, it was speculated that circNEK6 played an important role in the development of GCA via miR-532-5p、miR-339-5p、miR-665/HNF4A/NEK6/Cyclin D2 axis. This study aims to investigate the mechanisms by which circNEK6 promotes the carcinogenesis of GCA through gain and loss experiments, fluorescence in situ hybridization, ChIRP-Seq, PAR-Clip methods etc, so as to provide new preventive and therapeutic strategies for gastric cardiac adenocarcinoma.

本课题组前期在国际上率先发现：circNEK6在贲门腺癌组织中的表达相对癌旁正常组织显著上调，下调其表达可显著抑制HNF4A、NEK6及Cyclin D2表达并上调miR-532-5p、miR-339-5p、miR-665表达，进而抑制贲门腺癌细胞克隆形成。而且，NEK6为circNEK6宿主基因，HNF4A、NEK6及Cyclin D2为miR-532-5p、miR-339-5p、miR-665靶基因，HNF4A亦是NEK6及Cyclin D2的潜在转录因子。因此推测: circNEK6通过miR-532-5p、miR-339-5p、miR-665/HNF4A/NEK6/Cyclin D2通路促进贲门腺癌形成。本项目拟利用Gain-及Loss-实验、荧光原位杂交、ChIRP-Seq、PAR-Clip等方法进一步探讨circNEK6促进贲门腺癌形成的分子机制，以期为贲门腺癌防治提供新策略。

本课题组前期发现：circNEK6在胃贲门腺癌组织中的表达相对癌旁正常组织显著上调，下调其表达可显著下调HNF4A、NEK6及Cyclin D2表达并上调miR-532-5p、miR-339-5p、miR-665表达，并抑制贲门腺癌细胞克隆形成。而且，NEK6为circNEK6宿主基因，HNF4A、NEK6及Cyclin D2为miR-532-5p、miR-339-5p、miR-665靶基因，HNF4A亦是NEK6及Cyclin D2的潜在转录因子。因此推测: circNEK6通过miR-532-5p、miR-339-5p、miR-665/HNF4A/NEK6/Cyclin D2通路促进贲门腺癌形成。本研究对于研究circNEK6/miR-532-5p、miR-339-5p、miR-665/HNF4A/NEK6/Cyclin D2信号通路在胃贲门腺癌发生发展中分子机制的理解并寻找胃癌潜在治疗靶点具有重要意义。在本项目支持下发表SCI论文3篇，培养硕士2名，均已取得硕士学位，项目投入经费45万元，支出40.4万元，各项支出无超出预算情况，剩余经费4.5万余元，剩余经费计划用于本项目研究后续支出。