Hippo信号对卵巢生殖干细胞增殖及卵巢功能的影响

Effect of Hippo Signaling on Proliferation of Ovarian Reproductive Stem Cells and Ovarian Function

已有研究表明,Hippo信号通路对干细胞的自我更新和分化至关重要,且Hippo信号通路在调控卵泡生长中起重要作用,然而,目前关于Hippo通路对卵巢生殖干细胞的增殖和分化以及卵巢功能重塑的影响相关的研究较少。为了明确Hippo信号通路效应因子YAP1与卵巢生殖干细胞体外增殖分化的关系,以及Hippo信号通路对卵巢癌的主要功能。我们采用两步法酶促分离和磁性分离技术分别鉴定卵巢生殖干细胞,通过测定MVH和OCT4标记物的表达,然后选择YAP1作为Hippo信号通路的主要效应分子,作为研究的靶基因。将含有过表达的YAP1或YAP1靶向的shRNA的慢病毒转导入卵巢生殖干细胞中。通过将过表达YAP1或YAP1 shRNA的慢病毒载体微量注射到不育小鼠模型中,观察调节Hippo信号通路对卵巢的增殖、分化和内分泌功能的影响。研究结果表明,在分离的卵巢生殖干细胞中观察到YAP1和MVH的共表达。与对照组相比,过表达YAP1的卵巢生殖干细胞中MVH和OCT4表达水平显著增加。而YAP1敲低后,MVH和OCT4水平显著降低;不育小鼠模型中YAP1过表达15 d后,E2和FSH含量显著升高,而YAP1 shRNA表达后,小鼠血清E2和FSH含量显著降低。YAP1可用于调控卵巢生殖干细胞的增殖和分化以及小鼠的卵巢功能。本研究表明,Hippo信号通路可能是调控卵巢功能重建的一个新的分子靶点。

Previous studies have shown that Hippo signaling pathway is essential for self-renewal and differentiation of stem cells, and it plays an important role in regulating follicular growth. However, there are few studies on the effect of Hippo pathway on proliferation and differentiation of ovarian reproductive stem cells and remodeling of ovarian function. In order to clarify the relationship between Hippo signaling pathway effector YAP1 and ovarian germline stem cell proliferation and differentiation in vitro, and the main function of Hippo signaling pathway in ovarian cancer, two-step enzymatic separation and magnetic separation were used to identify ovarian germ cells and determine the expression of MVH and OCT4 markers respectively, while YAP1 was selected as the main effector of the Hippo signaling pathway and as the target gene for the study. Lentiviruses containing overexpressed YAP1 or YAP1 targeted shRNA were transduced into ovarian germline stem cells. The effects of Hippo signaling pathway on ovarian proliferation, differentiation and endocrine function were observed by microinjection of lentiviral vector overexpressing YAP1 or YAP1 shRNA into infertility mouse model. The results showed that co-expression of YAP1 and MVH was observed in isolated ovarian germline stem cells.Compared with the control group, the expression levels of MVH and OCT4 increased significantly in ovarian germ cells after YAP1 overexpression. However, the levels of MVH and OCT4 decreased significantly after YAP1 knockdown. The levels of E2 and FSH in infertile mice increased significantly after 15 days of YAP1 overexpression, while the levels of E2 and FSH decreased significantly after YAP1 shRNA expression. YAP1 can be used to regulate ovarian germline stem cell proliferation and differentiation and ovarian function in mice. This study indicated that Hippo signaling pathway may be a new molecular target for regulating ovarian function reconstruction.