Fang Fang, Li Dawei, Zhao Lu, Li Yue, Zhang Teng, Cui Baoxia
Department of Obstetrics and Gynecology, Qilu Hospital, Shandong University, Jinan, Shandong, People's Republic of China,
Department of Obstetrics and Gynecology, Weihai Municipal Hospital, Weihai, Shandong, People's Republic of China.
Onco Targets Ther. 2019 Jan 18;12:685-697. doi: 10.2147/OTT.S180534. eCollection 2019.
Our study aimed to investigate the expression of NR1H3 in endometrial carcinoma, its effect on the proliferation of endometrial carcinoma cells in vitro, and the underlying mechanism of this effect.
Immunohistochemistry of paraffin-embedded, sectioned specimens and of a tissue microarray was conducted to estimate the expression of NR1H3 (liver X receptors α: LXRα) and NR1H2 (liver X receptors β: LXRβ) in endometrial carcinoma tissues. The subcellular localization of NR1H3 in the endometrial carcinoma cell line Ishikawa was determined by immunofluorescence. An agonist of NR1H3, TO901317, was then administered to activate the expression of NR1H3, and cell viability and cell-cycle progression were investigated through MTT and flow cytometric assays, respectively. The gene and protein expression levels of NR1H3, cyclin D1 (CCND1), and cyclin E (CCNE) in cells pretreated with different concentrations of TO901317 for different periods of time were also detected by real-time RT-PCR and Western blot, respectively.
The results showed that, in contrast to NR1H2, which was expressed at low levels in endometrial tissues, NR1H3 was upregulated in endometrial adenocarcinoma tissues compared to levels in normal endometrial tissues and endometrial polyps. Moreover, NR1H3 was mainly expressed in the cytoplasm of Ishikawa cells. TO901317 significantly decreased cell viability and arrested the cell cycle in Ishikawa cells in a dose- and time-dependent manner. Furthermore, the administration of TO901317 not only promoted the expression of NR1H3 but also inhibited the expression of CCND1 and CCNE in Ishikawa cells.
We demonstrated that NR1H3 is upregulated in endometrial adenocarcinoma and that it inhibits cell viability by inhibiting the expression of CCND1 and CCNE in endometrial carcinoma cells. Our study indicates that NR1H3 may play a role in the development of endometrial cancer and may emerge as a promising therapeutic target.
本研究旨在探讨NR1H3在子宫内膜癌中的表达情况、其对子宫内膜癌细胞体外增殖的影响以及这种影响的潜在机制。
对石蜡包埋、切片的标本以及组织芯片进行免疫组织化学检测，以评估NR1H3（肝脏X受体α：LXRα）和NR1H2（肝脏X受体β：LXRβ）在子宫内膜癌组织中的表达。通过免疫荧光法确定NR1H3在子宫内膜癌细胞系Ishikawa中的亚细胞定位。然后给予NR1H3激动剂TO901317以激活NR1H3的表达，并分别通过MTT法和流式细胞术检测细胞活力和细胞周期进程。还分别通过实时RT-PCR和蛋白质免疫印迹法检测了在不同时间段用不同浓度TO90131预处理的细胞中NR1H3、细胞周期蛋白D1（CCND1）和细胞周期蛋白E（CCNE）的基因和蛋白表达水平。
结果显示，与在子宫内膜组织中低表达的NR1H2不同，与正常子宫内膜组织和子宫内膜息肉中的水平相比，NR1H3在子宫内膜腺癌组织中上调。此外，NR1H3主要在Ishikawa细胞的细胞质中表达。TO901317以剂量和时间依赖性方式显著降低Ishikawa细胞的活力并使细胞周期停滞。此外，给予TO901317不仅促进了NR1H3的表达，还抑制了Ishikawa细胞中CCND1和CCNE的表达。
我们证明NR1H3在子宫内膜腺癌中上调，并且它通过抑制子宫内膜癌细胞中CCND1和CCNE的表达来抑制细胞活力。我们的研究表明NR1H3可能在子宫内膜癌的发生发展中起作用，并可能成为一个有前景的治疗靶点。
