miR-361-3P调控转化生长因子β1诱导成骨细胞凋亡的研究  

Effect of miR-361-3p on TGF-β1-induced apoptosis in osteoblasts

作　　者：王帆[1] 王鹏皓 WANG Fan;WANG Penghao(Department of Orthopaedics,The First Hospital of China Medical University,Shenyang 110001,China)

机构地区：[1]中国医科大学附属第一医院骨科,沈阳110001

出　　处：《中国医科大学学报》2023年第7期577-581,共5页Journal of China Medical University

基　　金：国家自然科学基金(81971322)。

摘　　要：目的研究人成骨细胞内miR-361-3P对转化生长因子β1(TGF-β1)和细胞凋亡标志分子caspase 3蛋白活性表达的影响,探讨miR-361-3P/TGF-β1通路在人成骨细胞凋亡中的作用机制。方法用miR-361-3p mimic或inhibitor及TGF-β1 siRNA(siTGF-β1)转染人成骨细胞系hFob1.19;RT-qPCR和Western blotting检测caspase 3的变化;CCK-8法检测细胞的增殖和凋亡;荧光素酶基因报告实验检测miR-361-3p和TGF-β1基因的结合情况。结果在hFob1.19细胞内,miR-361-3p mimic显著下调了TGF-β1mRNA表达,同时明显抑制了细胞内的TGF-β1蛋白活性(P<0.05);miR-361-3p mimic作用hFob1.19细胞48 h后,miR-361-3p可明显抑制细胞增殖(P<0.05),导致成骨细胞内caspase 3蛋白活性明显升高(P<0.05),并减少WT-TGF-β1的荧光素酶活性;miR-361-3p inhibitor可增强hFob1.19细胞增殖,抑制细胞内caspase 3蛋白活性,而si-TGF-β1则可明显增强caspase 3活性表达。结论miR-361-3p可能通过抑制TGF-β1负向调控人成骨细胞的增殖并促进其凋亡。Objective To investigate the effect of microRNA(miR)-361-3p on transforming growth factor-β1(TGF-β1)expression and apoptosis marker molecule caspase 3 protein activity,and the apoptotic mechanism of the miR-361-3p/TGF-1 pathway in human osteoblasts.Methods The miR-361-3p mimic or miR-361-3p and small interferingRNA(si-TGF-β1)were transfected into human osteoblasts(hFob1.19).The expression and binding of miR-361-3p and TGF-β1 were assessed by real-time quantitative PCR(RT-qPCR),CCK-8 kit,and luciferase gene reporter assay.Results Within hFob1.19 cells,the miR-361-3p mimic significantly downregulated TGF-1 mRNA expression and significantly inhibited TGF-β1 protein activity(P<0.05).Cell proliferation was significantly decreased in hFob1.19 cells after 48 h by the miR-361-3p mimic(P<0.05),and the activity of caspase 3 significantly increased(P<0.05).The miR-361-3p mimic also reduced the luciferase activity of WT-TGF-β1.The miR-361-3p inhibitor enhanced the proliferation of hFob1.19 cells and inhibited the protein activity of caspase 3 in the cells,while si-TGF-β1 enhanced the active expression of caspase 3.Conclusion miR-361-3p may negatively regulate the proliferation of hFob1.19 cells and promote apoptosis by inhibiting TGF-β1.

关 键 词：成骨细胞 miR-361-3P 转化生长因子Β1 caspase 3 细胞凋亡 

分 类 号：R336[医药卫生—人体生理学]