TBL1XR1通过上调Wnt/β-catenin信号通路促进卵巢癌细胞迁移和侵袭

TBL1XR1 promotes the migration and invasion of ovarian cancer cells by upregulating Wnt/β-catenin signaling pathway

目的探讨转导蛋白(β)样1 X连锁受体1(TBL1XR1)通过上调Wnt/β-catenin信号通路促进卵巢癌细胞迁移和侵袭的机制。方法使用基因表达谱交互分析工具(GEPIA 2.0)、免疫组化和Western blotting评估TBL1XR1在卵巢癌和正常卵巢组织中的差异表达;评估TCGA数据库中TBL1XR1与卵巢癌患者总生存期的相关性,对TBL1XR1高、低表达组之间的差异表达基因进行GO和KEGG富集分析;分别采用伤口愈合实验、Transwell基质胶侵袭实验检测TBL1XR1对卵巢癌细胞迁移和侵袭能力的影响;Western blotting检测敲低或过表达TBL1XR1后Wnt/β-catenin信号通路蛋白表达水平的变化;TOP/FOP荧光素酶报告基因检测TBL1XR1对Wnt/β-catenin信号通路活性变化的影响。结果卵巢癌组织中TBL1XR1表达水平高于正常卵巢组织,低表达组卵巢癌患者总生存期长于高表达组;GO和KEGG分析结果提示,TBL1XR1的生物学功能与激素转运、激素分泌、激素分泌的调节及肽转运、神经活性配体-受体相互作用相关;敲低TBL1XR1可显著抑制卵巢癌细胞的迁移和侵袭,且Wnt3a能逆转这一效应,过表达TBL1XR1可显著提高卵巢癌细胞迁移和侵袭能力;下调TBL1XR1表达可显著降低Wnt/β-catenin信号通路的活性及蛋白表达水平。结论TBL1XR1在卵巢癌中高表达且与卵巢癌患者不良预后相关,TBL1XR1能够通过上调Wnt/β-catenin信号通路促进卵巢癌细胞的迁移和侵袭,可能作为促癌因子在卵巢癌的发生发展中发挥重要作用。

Objective To investigate the mechanism of transducin(β)-like 1 X-linked receptor 1(TBL1XR1)promoting the migration and invasion of ovarian cancer cell by upregulating the Wnt/β-catenin signaling pathway.Methods Differen-tial expression of TBL1XR1 in ovarian cancer and normal ovarian tissues was assessed by using the Gene Expression Profil-ing Interactive Analysis(GEPIA 2),immunohistochemistry and Western blotting methods.The correlation between TBL1XR1 and overall survival of ovarian cancer patients in the Cancer Genome Atlas(TCGA)database was evaluated.The GO and KEGG enrichment analysis of differentially expressed genes(DEG)between TBL1XR1 high and low expression groups were also performed to explore their bioinformatic functions.Wound healing assay and Transwell stromal gel inva-sion assay were used to determine the effect of TBL1XR1 on the migration and invasion ability of ovarian cancer cells,re-spectively.Rescue experiments were performed with Wnt3a,an activator of the Wnt/β-catenin signaling pathway.Western blotting was used to detect changes in Wnt/β-catenin signaling pathway protein expression levels after knockdown or over-expression of TBL1XR1.The effect of TBL1XR1 on the changes of Wnt/β-catenin signaling pathway activity was detected by TOP/FOP luciferase reporter assay.Results The expression level of TBL1XR1 in ovarian cancer tissues was higher than that in normal ovarian tissues,and the overall survival of ovarian cancer patients in the low expression group was higher than that in the high expression group.The results of GO analysis and KEGG analysis suggested that the biological func-tions of TBL1XR1 were related to hormone transport,hormone secretion,regulation of hormone secretion and peptide trans-port,and neuroactive ligand-receptor interactions.Knockdown of TBL1XR1 significantly inhibited the migration and inva-sion of ovarian cancer cell,and this effect could be reversed by Wnt3a.Overexpression of TBL1XR1 significantly increased the migration and invasion ability of ovarian cancer cells.When TBL1XR1 expression was downregulated,the activity of Wnt/β-catenin signaling pathway and its protein expression levels were subsequently reduced.Conclusion TBL1XR1 was highly expressed in ovarian cancer and was associated with the poor prognosis of ovarian cancer patients.TBL1XR1 could promote the migration and invasion of ovarian cancer cells by upregulating Wnt/β-catenin signaling pathway.It may play an important role in the tumorigenesis and development of ovarian cancer as a tumor promoter.