Zhang Peng, Li Jifeng, Song Yuze, Wang Xiao
Cell Physiol Biochem. 2017;42(1):242-253. doi: 10.1159/000477323. Epub 2017 May 25.
BACKGROUND/AIMS: Recently, microRNAs (miRNA) have been identified as novel regulators in Chondrosarcoma (CHS). This study was aimed to identify the roles of miR-129-5p-5p in regulation of SOX4 and Wnt/β-catenin signaling pathway, as well as cell proliferation and apoptosis in chondrosarcomas.
Tissue samples were obtained from chondrosarcoma patients. Immunohistochemistry, real-time quantitative RT-PCR (RT-qPCR) and western blot analysis were performed to detect the expressions of miR-129-5p and SOX4. Luciferase assay was conducted to confirm that miR-129-5p directly targeted SOX4 mRNA. Manipulations of miR-129-5p and SOX4 expression were achieved through cell transfection. Cell proliferation, migration and apoptosis were evaluated by CCK-8 assay, colony forming assay, wound healing assay and flow cytometry in vitro. For in vivo experiment, the tumor xenograft model was established to evaluate the effects of miR-129-5p and SOX4 on chondrosarcomas.
The expression of miR-129-5p was significantly down-regulated in chondrosarcoma tissues as well as cells in comparison with normal ones, while SOX4 was over-activated. Further studies suggested that miR-129-5p suppressed cell proliferation, migration and promoted apoptosis by inhibiting SOX4 and Wnt/β-catenin pathway.
MiR-129-5p inhibits the Wnt/β-catenin signaling pathway by targeting SOX4 and further suppresses cell proliferation, migration and promotes apoptosis in chondrosarcomas.
背景/目的:最近,微小RNA(miRNA)已被确定为软骨肉瘤(CHS)中的新型调节因子。本研究旨在确定miR-129-5p在调节SOX4和Wnt/β-连环蛋白信号通路以及软骨肉瘤细胞增殖和凋亡中的作用。
从软骨肉瘤患者获取组织样本。进行免疫组织化学、实时定量逆转录聚合酶链反应(RT-qPCR)和蛋白质印迹分析以检测miR-129-5p和SOX4的表达。进行荧光素酶测定以证实miR-129-5p直接靶向SOX4 mRNA。通过细胞转染实现对miR-129-5p和SOX4表达的调控。在体外通过CCK-8测定、集落形成测定、伤口愈合测定和流式细胞术评估细胞增殖、迁移和凋亡。对于体内实验,建立肿瘤异种移植模型以评估miR-129-5p和SOX4对软骨肉瘤的影响。
与正常组织和细胞相比,软骨肉瘤组织及细胞中miR-129-5p的表达显著下调,而SOX4过度激活。进一步研究表明,miR-129-5p通过抑制SOX4和Wnt/β-连环蛋白途径抑制细胞增殖、迁移并促进凋亡。
MiR-129-5p通过靶向SOX4抑制Wnt/β-连环蛋白信号通路,并进一步抑制软骨肉瘤细胞的增殖、迁移并促进其凋亡。