LncRNA APTR促进雌激素调控ERα/Wnt通路增强子宫肌瘤细胞增殖的机制研究

Uterine leiomyoama is caused by abnormal proliferation of uterine smooth muscle. Elucidating the mechanism of abnormal prolieration of uterine leiomyoma cells is the focus of current study. The proliferation of uterine fibroids is estrogen dependent, however, the molecular mechanism of estrogen regulating the proliferation of uterine leiomyoma is still not clear. Our previous study found that the regulation of estrogen on the proliferation of uterine leiomyoma cells is mediated by ERα receptor. ERα regulates the proliferation of the uterine leiomyoma by regulating the expression of key factors of Wnt signaling pathway. By highthroughput sequencing, LncRNA APTR is highly expressed in uterine leiomyoma tissue, and directly binds the complex of ERα/Sp1, then regulating the activity of Wnt signaling pathway leading to regulation of the proliferation of uterine fibroids. In this sutdy, we will clearify the role of LncRNA-APTR-ER signaling pathway by molecular biology, biochemistry, model animal in vivo experiment, clinical specimens and in vitro cell experiment mathods. The implementation of this project will lay a theoretical foundation for elucidating the pathogenesis of uterine fibroids and exploring new treatment methods.

子宫肌瘤是由子宫平滑肌异常增殖造成的，阐明子宫肌瘤细胞异常增殖的调控机制至关重要。子宫肌瘤的增殖是雌激素依赖性的，但是雌激素调控子宫肌瘤细胞增殖的分子机制尚不明晰。我们前期研究发现：雌激素对子宫肌瘤细胞增殖的调控作用由ERα介导的；ERα通过调控Wnt信号通路关键因子的表达进而调控子宫肌瘤增殖。更有意义的是：通过高通量测序检测子宫肌瘤特异性转录组，我们发现LncRNA APTR在子宫肌瘤组织中高度特异性表达；生物信息学分析和初步生化实验结果提示LncRNA APTR通过直接结合ERα/Sp1复合物，调控Wnt信号通路活性，从而调控子宫肌瘤的增殖。在本项目中，我们将结合分子生物学、生物化学、模式动物体内实验、子宫肌瘤临床标本和体外细胞实验等手段明确LncRNA APTR-ER通路对子宫肌瘤的调控作用，阐明其作用机理。本项目的实现将为阐明子宫肌瘤发病机理，探索新的治疗方法奠定理论基础。

子宫肌瘤是最常见的妇科良性肿瘤，影响患者的生活质量，是临床子宫切除的首要原因。近年来，女性生活习惯改变导致婚育年龄增高，鉴于生育的要求，临床迫切需求控制肿瘤生长的非手术治疗方法，安全、有效的保守性治疗方法成为目前研究的热点。长链非编码RNA（Long non-coding RNA, LncRNA）是长度大于 200 个核苷酸的非编码 RNA，参与调控细胞增殖等多项生物学行为。申请人课题组首次发现LncRNA-ENST00000447009 (LncRNA APTR) 在子宫肌瘤中特异性高表达，并且促进子宫肌瘤细胞增殖。此项研究延续以前的工作并进一步证实：成功分离并鉴定原代子宫平滑肌细胞，进一步证实APTR促进子宫肌瘤细胞增殖，ERa是APTR促进子宫肌瘤细胞增殖的靶点，APTR通过调控Wnt通路促进子宫肌瘤细胞增殖。该研究成果发表在期刊Frontiers in Oncology。另外我们还发现子宫肌瘤细胞中Wnt16呈高表达，雌激素通过ERα增高Wnt16的表达，Wnt16促进子宫肌瘤细胞增殖，且通过Wnt/Ca+通路促进子宫肌瘤细胞增殖，研究成果在投稿中。该项研究从理论上丰富了ERα上游和下游的调控靶点，完善了雌激素促进子宫肌瘤细胞增殖的分子机制，项目的顺利完成，为子宫肌瘤保守性治疗提供了新的靶点，为子宫肌瘤非手术治疗方法带来了曙光。该项目的完成具有明显潜在的社会经济效益和应用前景。