miR-98-5p促进成骨细胞增殖和分化的可能及机制  

MicroRNA-98-5p promotes osteoblast proliferation and differentiation:possibilities and mechanisms

作　　者：郑峰[1] 张富财[1] 许喆[1] Zheng Feng;Zhang Fucai;Xu Zhe(Department of Orthopedics,Qinghai Provincial People’s Hospital,Xining 810007,Qinghai Province,China)

机构地区：[1]青海省人民医院骨科,青海省西宁市810007

出　　处：《中国组织工程研究》2021年第26期4112-4117,共6页Chinese Journal of Tissue Engineering Research

基　　金：青海省基础研究计划项目(2020-ZJ-755),项目负责人:郑峰。

摘　　要：背景:miRNA-98-5p可抑制高迁移率族AT HOOK蛋白2(high mobility group AT-HOOK 2,HMGA2)。磷脂酰肌醇3-激酶/蛋白激酶B/糖原合成酶激酶3β(phosphatidylinositol 3-kinase/alkaline phosphatase/glycogen synthase kinase-3β,PI3K/AKT/GSK-3β)信号通路参与细胞增殖过程。骨折愈合过程中,miR-98-5p是否通过激活HMGA2作用于PI3K/Akt/GSK-3β通路来促进成骨细胞的增殖仍不清楚。目的:探讨miR-98-5p调控HMGA2和PI3K/AKT/GSK-3β通路促进成骨细胞增殖和分化的机制。方法:取MC3T3-E1细胞,通过Lipofectamine 2000分别转染miR-98-5p模拟物和HMGA2质粒到MC3T3-E1细胞,分别为miR-98-5p转染组和HMGA2过表达组;用二甲基亚砜处理的MC3T3-E1细胞及Scramblez乱序质粒转染至MC3T3-E1,分别为转染对照组和空白质粒组;采用脂质体瞬时转染miR-98-5p抑制剂,作为miR-98-5p抑制剂组;不做任何处理的MC3T3-E1细胞作为空白对照组。结果与结论:①空白对照组和转染对照组的HMGA2、PI3K/Akt/GSK-3β、碱性磷酸酶活性及mRNA水平差异无显著性意义,与转染对照组比,miR-98-5p转染组HMGA2和PI3K/Akt/GSK-3β蛋白表达、细胞分化、细胞增殖,碱性磷酸酶活性及mRNA均显著降低;②Targetscan7.1预测miR-98-5p与HMGA2相互作用结果显示,与转染对照组比,miR-98-5p转染组的HMGA2蛋白和mRNA均降低;与miR-98-5p转染组比,miR-98-5p抑制剂组的HMGA2蛋白和mRNA均增加;③空白对照组和空白质粒组的成骨细胞增殖、碱性磷酸酶活性及mRNA水平差异无显著性意义,与空白质粒组比,HMGA2过表达组的PI3K/Akt/GSK-3β表达、细胞分化程度、MC3T3-E1细胞增殖、碱性磷酸酶活性及mRNA水平均显著增加;④结果显示miR-98-5p可抑制MC3T3-E1细胞的HMGA2和PI3K/Akt/GSK-3β表达,同时抑制细胞的增殖和分化,HMGA2可能是miR-98-5p的直接作用靶点。BACKGROUND:MicroRNA-98-5p(miR-98-5p)can inhibit the high mobility group AT-HOOK 2(HMGA2).The phosphatidylinositol 3-kinase/alkaline phosphatase/glycogen synthase kinase-3β(PI3K/AKT/GSK-3β)signaling pathway is involved in cell proliferation.During the fracture healing process,it is unclear whether miR-98-5p can act on the PI3K/Akt/GSK-3βpathway by activating HMGA2 to promote the proliferation of osteoblasts.OBJECTIVE:To investigate the mechanisms of microRNA-98-5p(miR-98-5p)in promoting osteoblast proliferation and differentiation by regulating HMGA2 and PI3K/Akt/GSK-3βpathway.METHODS:MC3T3-E1 cells transfected with miR-98-5p mimics and HMGA2 plasmid by Lipofectamine 2000 were divided into a miR-98-5p mimics group and a HMGA2 overexpression group,respectively.MC3T3-E1 cells transfected with dimethyl sulfoxide and scrambled plasmids were divided into a mimic control group and a scramble group.Cells transiently transfected with miR-98-5p inhibitor using liposomes were as a miR-98-5p inhibitor group.Normal MC3T3-E1 cells without treatment were used as a blank control group.RESULTS AND CONCLUSION:There were no significant differences in HMGA2,PI3K/Akt/GSK-3β,alkaline phosphatase activity and mRNA level between the blank control group and the mimic control group.Compared with the mimic control group,the HMGA2 and PI3K/Akt/GSK-3βprotein expression,cell differentiation and proliferation,alkaline phosphatase activity and mRNA level were significantly reduced in the miR-98-5p mimics group.The interaction between miR-98-5p and HMGA2 predicted by Targetscan7.1 showed that compared with the mimic control group,the HMGA2 protein and mRNA were reduced in the miR-98-5p mimic group,while compared with the miR-98-5p mimic group,the HMGA2 protein and mRNA had an increase in the miR-98-5p inhibitor group.There were no significant differences in osteoblast proliferation,alkaline phosphatase activity and mRNA between the blank control group and the scramble group.Compared with the scramble group,the PI3K/Akt/GSK-3βexpression,ce

关 键 词：成骨细胞 小微RNA98-5p HMGA2 磷脂酰肌醇3-激酶 蛋白激酶B 糖原合成酶激酶3Β 转染 MC3T3-E1细胞 增殖 分化 

分 类 号：R446[医药卫生—诊断学] R363[医药卫生—临床医学]