反义长链非编码RNA HHIP-AS1调控hedgehog信号通路影响子宫内膜癌发生发展的机制研究

The incidence for endometrial cancer is on rise in the past a few years, and it becomes a serious threat to women’s health in China. Previous evidence has identified long non-coding RNA(LncRNA) to be closely associated with cancer occurrence and development, however, the role of LncRNA on endometrial cancer remains unclearly decoded. By using the RNA-sequncing platform, we have previously identified a most significantly down-expression antisense LncRNA HHIP-AS1 in endometrial cancer patients. The previous experiments also showed that the HHIP-AS1 was closely in relevance with down-expression of the HHIP gene which is a core negative regulatory factor for hedgehog pathway associated with endometrial carcinogenesis. Further bioinformatics analysis indicated that there are binding sites of HHIP-AS1 at the promoter region of HHIP. Thus, we hypothesized that the down-expression of HHIP-AS1 is associated with endometrial cancer initiation and progression via its regulation on HHIP and hedgehog signaling pathway. Here, the project intends to estimate the association between the HHIP-AS1 and endometrial cancer risk as well as prognosis by detecting the antisense LncRNA expression in large clinal samples. A series of functional assays will be used to illustrate the biological mechanism of HHIP-AS1 on regulating hedgehog signaling pathway in endometrial cancer development. This project will help us to decipher the mechanism of endometrial cancer pathology, and provide scientific evidences for targeted therapy of endometrial cancer.

子宫内膜癌发病率逐年上升，已严重危害我国女性的生命健康。长链非编码RNA（LncRNA）与肿瘤发生发展关系密切，但有关子宫内膜癌的LncRNA研究尚处于起步阶段。课题组基于RNA-seq预实验发现反义LncRNA HHIP-AS1在子宫内膜癌癌组织中低表达，且与hedgehog信号通路核心负调控因子HHIP表达呈正相关。生物信息学分析还发现HHIP基因启动子存在该反义LncRNA的结合序列。hedgehog信号通路是影响子宫内膜癌变的关键通路，故推测HHIP-AS1可通过调控HHIP基因调节hedgehog信号通路而影响子宫内膜癌的发生发展。本项目拟在前期研究基础上，进一步探讨HHIP-AS1表达与子宫内膜癌发病、进展及预后的关系；通过表型检测和一系列体内、外功能实验探讨该反义LncRNA调控hedgehog信号通路的生物学机制，为子宫内膜癌的靶向治疗提供依据。

非编码RNA在恶性肿瘤发生发展过程起着关键的作用。解析其在肿瘤进程中的相关调控网络及功能机制，并应用于肿瘤防治具有重要的现实意义。本研究旨在预试验筛查结果的基础上，分析了长链非编码RNA HHIP-AS1与我国人群子宫内膜癌发生发展的关联及其生物学机制。利用qRT-PCR检测了 HHIP-AS1在子宫内膜癌及癌旁组织的表达情况,发现HHIP-AS1在癌组织中的表达水平显著低于对应的癌旁正常组织。HHIP-AS1表达与子宫内膜癌临床分期密切相关，低表达HHIP-AS1能显著促进内膜癌患者的临床进展。系列功能实验显示，过表达HHIP-AS1可显著抑制子宫内膜癌细胞系的增殖、克隆形成、细胞迁移和侵袭、裸鼠成瘤能力。HHIP-AS1可通过竞争性结合miR-96而调控抑癌基因HHIP的表达,进而影响下游Hedgehog信号通路相关基因的表达水平。以上结果说明，HHIP-AS1可通过调控Hedgehog信号通路而影响我国人群子宫内膜癌的发生发展。本研究为子宫内膜癌的基础研究和临床靶向治疗提供了新思路和分子靶标。