Abstract：
To preliminarily explore miR-503 in human degenerative disc nucleus pulposus cell effects as well as mechanisms.
We utilized bioinformatics analysis to determine the miRNA differential expression as well as key signal pathways existing in human nucleus pulposus cells of the degenerative intervertebral discs. Human degenerative disc nucleus pulposus cell model was cultured and established in vitro. miR-503 and TNIK-related genes are knocked down and overexpressed by lentiviral infection, then we added Wnt signaling pathway agonists; CCK-8, ELISA, RT-PCR, Western blot were used to detect proliferation, apoptosis, and activity of cells.
Bioinformatics results demonstrated that miR-503 was significantly down-regulated in human nucleus pulposus cells of the degenerated intervertebral discs. The targeted differentially expressed genes were mainly enriched in Wnt signaling pathway. However, after screening differential genes in the Wnt pathway, it was demonstrated that miR-503 mainly regulates TNIK to achieve Wnt pathway regulation. Cell experiments in vitro showed that cell activity and function were decreased while apoptosis was increased in the degenerative cell model.
miR-503 can improve the function and activity of human nucleus pulposus cells of degenerated intervertebral disc by inhibiting Wnt expression. miR-503 mainly regulates the Wnt pathway through targeted binding with TNIK.
We utilized bioinformatics analysis to determine the miRNA differential expression as well as key signal pathways existing in human nucleus pulposus cells of the degenerative intervertebral discs. Human degenerative disc nucleus pulposus cell model was cultured and established in vitro. miR-503 and TNIK-related genes are knocked down and overexpressed by lentiviral infection, then we added Wnt signaling pathway agonists; CCK-8, ELISA, RT-PCR, Western blot were used to detect proliferation, apoptosis, and activity of cells.
Bioinformatics results demonstrated that miR-503 was significantly down-regulated in human nucleus pulposus cells of the degenerated intervertebral discs. The targeted differentially expressed genes were mainly enriched in Wnt signaling pathway. However, after screening differential genes in the Wnt pathway, it was demonstrated that miR-503 mainly regulates TNIK to achieve Wnt pathway regulation. Cell experiments in vitro showed that cell activity and function were decreased while apoptosis was increased in the degenerative cell model.
miR-503 can improve the function and activity of human nucleus pulposus cells of degenerated intervertebral disc by inhibiting Wnt expression. miR-503 mainly regulates the Wnt pathway through targeted binding with TNIK.
摘要：
目的：初步探讨miR-503在人退变椎间盘髓核细胞中的作用及机制。
方法：我们利用生物信息学分析来确定人椎间盘退变髓核细胞中存在的miRNA差异表达以及关键信号通路。体外培养并建立人椎间盘退变髓核细胞模型。miR-503和TNIK相关基因被慢病毒感染敲低并过表达，然后我们加入Wnt信号通路激动剂；CCK-8，ELISA，RT-PCR,Westernblot用于检测增殖，凋亡，和细胞的活动。
结果：生物信息学结果表明，miR-503在退变椎间盘的人髓核细胞中显著下调。靶向的差异表达基因主要富集在Wnt信号通路中。然而,在筛选Wnt途径中的差异基因后，已证明miR-503主要调节TNIK以实现Wnt通路调节。体外细胞实验表明，在变性细胞模型中，细胞活性和功能降低，而凋亡增加。
结论：miR-503可通过抑制Wnt的表达提高退变椎间盘髓核细胞的功能和活性。miR-503主要通过与TNIK的靶向结合来调节Wnt途径。
方法：我们利用生物信息学分析来确定人椎间盘退变髓核细胞中存在的miRNA差异表达以及关键信号通路。
结果：生物信息学结果表明，miR-503在退变椎间盘的人髓核细胞中显著下调。
结论：miR-503可通过抑制Wnt的表达提高退变椎间盘髓核细胞的功能和活性。
