Mesenchymal-to-epithelial transition of osteoblasts induced by Fam20c knockout
Mesenchymal-to-epithelial transition of osteoblasts induced by Fam20c knockout
复制标题
Fam20c 敲除诱导的成骨细胞间充质到上皮细胞的转变
DOI：
10.1007/s13258-021-01170-4
复制
发表时间：
2022-01-13
期刊：
影响因子：
2.1
通讯作者：
Li, Ying
中科院分区：
文献类型：
--
作者：
Geng, Ya-Wei;Zhang, Zhen;Li, Ying
背景 Fam20c 与组织发育和疾病密切相关。目前已有报道Fam20c调节成骨细胞矿化，但其他作用的报道较少。目的通过敲除Fam20c基因，研究Fam20c对成骨细胞的影响。方法通过慢病毒转染小鼠成骨细胞，构建Fam20c基因敲除的成骨细胞。采用CCK-8、划痕试验和茜素红染色法检测Fam20c敲除细胞的增殖、迁移和矿化情况。通过透射电子显微镜观察亚细胞结构。采用RT-PCR检测间充质上皮转化（MET）相关标记基因和核心转录因子的差异表达。通过免疫荧光或Western blot检测MET相关蛋白的差异表达。对Fam20c敲除成骨细胞进行转录组分析，并使用实时PCR验证与MET相关的转录组分析。结果 Fam20c缺失后成骨细胞的增殖能力没有明显改变，但迁移能力和矿化能力明显减弱。 Fam20c 敲除细胞之间存在紧密连接。间充质细胞标志基因和核心转录因子的表达量显着降低，上皮细胞标志基因的表达量显着增加。间充质细胞标志蛋白表达显着降低，上皮细胞标志蛋白表达显着升高。 MET 涉及的多个信号分子和途径发生了变化。结论 Fam20c 的敲低导致 MET。 Fam20c 影响成骨细胞 MET 中关键因子的转录。
Background Fam20c is intimately related to tissue development and diseases. At present, it has been reported that Fam20c regulates the mineralization of osteoblasts, but there are few reports on other effects. Objective To study the effect of Fam20c on osteoblasts by knocking out the Fam20c gene. Methods Fam20c knockout osteoblasts were constructed by transfecting mouse osteoblasts with lentivirus. The proliferation, migration and mineralization of Fam20c knockout cells were detected by CCK-8, scratch test and alizarin red staining assays. The subcellular structure was observed by transmission electron microscopy. RT-PCR was used to detect the differential expression of mesenchymal-to-epithelial transition (MET)-related marker genes and core transcription factors. The differential expression of MET-related proteins was detected by immunofluorescence or Western blot. Transcriptome analysis of Fam20c knockout osteoblasts was performed, and real-time PCR was used to verify transcriptome analysis related to MET. Results The proliferation ability of osteoblasts was not significantly changed after Fam20c deletion, but the migration ability and mineralization ability were significantly weakened. There were tight junctions between Fam20c knockout cells. The expression of mesenchymal cell marker genes and core transcription factors was significantly decreased, and the expression of epithelial cell marker genes was significantly increased. The expression of mesenchymal cell marker proteins was significantly decreased, and the expression of epithelial cell marker proteins was significantly increased. Multiple signalling molecules and pathways involved in MET have changed. Conclusions Knockdown of Fam20c resulted in MET. Fam20c affects the transcription of key factors in osteoblast MET.