School of Biomedical Sciences and PharmacyUniversity of Newcastle, Callaghan, New South Wales, Australia.
Division of Molecular MedicineNSW Health Pathology, John Hunter Hospital, Newcastle, New South Wales, Australia.
Reproduction. 2018 Jan;155(1):61-71. doi: 10.1530/REP-17-0339. Epub 2017 Oct 24.
Recent studies showed that genetic aberrations in the gene, probably through the canonical WNT/β-catenin pathway, lead to the pathogenesis of uterine fibroids. However, a comprehensive analysis of the WNT pathway in -mutated and -wild-type fibroids has not been performed. The objective of this study was to determine the status of the WNT pathway in human fibroids. We performed Sanger sequencing to define the mutational status of fibroids and normal myometrium samples. qPCR arrays were carried out to determine the status of the WNT signaling pathway in -mutated and -wild-type fibroids. Liquid chromatography-mass spectrometry (LC-MS), Western blotting and immunohistochemistry were used to monitor the expression of β-catenin. We showed that β-catenin expression was increased in fibroids compared to the adjacent myometrium samples. However, β-catenin expression showed no correlation with mutation status. Of all the WNT signaling components, WNT inhibitors showed the greatest differences in expression between fibroids and controls. , a WNT inhibitor, was identified as the most significantly upregulated gene in fibroids. We cultured primary fibroid cells on hydrogels of known stiffness to decipher the influence of biomechanical cues on β-catenin expression and revealed increased levels of β-catenin when cells were cultured on a stiffer surface. In conclusion, our data showed that β-catenin expression in fibroids occurs independently of mutations. Biomechanical changes upregulate β-catenin expression in fibroids, providing an attractive avenue for developing new treatments for this disease.
最近的研究表明，基因的遗传异常，可能通过经典的 WNT/β-连环蛋白通路，导致子宫肌瘤的发病机制。然而，尚未对突变型和野生型纤维瘤中的 WNT 通路进行全面分析。本研究的目的是确定 WNT 通路在人类纤维瘤中的状态。我们通过桑格测序来确定纤维瘤和正常子宫肌层样本中的突变状态。进行 qPCR 阵列以确定突变型和野生型纤维瘤中 WNT 信号通路的状态。通过液相色谱-质谱（LC-MS）、Western blot 和免疫组织化学监测β-连环蛋白的表达。我们表明，与相邻的子宫肌层样本相比，β-连环蛋白在纤维瘤中的表达增加。然而，β-连环蛋白的表达与 突变状态无关。在所有 WNT 信号成分中，WNT 抑制剂在纤维瘤和对照之间的表达差异最大。作为一种 WNT 抑制剂，被鉴定为纤维瘤中表达上调最显著的基因。我们在已知硬度的水凝胶上培养原代纤维瘤细胞，以解析生物力学线索对β-连环蛋白表达的影响，并揭示出当细胞在更硬的表面上培养时，β-连环蛋白的水平增加。总之，我们的数据表明，纤维瘤中β-连环蛋白的表达与 突变无关。生物力学变化上调纤维瘤中的β-连环蛋白表达，为开发这种疾病的新治疗方法提供了一个有吸引力的途径。
