精氨酸甲基转移酶(PRMT)7通过IGF-1信号通路调控人骨髓间充质干细胞成脂分化的研究  

PRMT7 Regulates Adipogenic Differentiation of hBMSCs by Modulating IGF-1 Signaling

作　　者：郭倩[1] 卿佳 陆大壮 王叙 李扬[1] 张慧[1] 张应飞 刘云松 周永胜 张萍[1] GUO Qian;QING Jia;LU Da-Zhuang;WANG Xu;LI Yang;ZHANG Hui;ZHANG Ying-Fei;LIU Yun-Song;ZHOU Yong-Sheng;ZHANG Ping(Department of Prosthodontics,Peking University School and Hospital of Stomatology,Beijing Key Laboratory of Digital Stomatology,National Engineering Laboratory for Digital and Material Technology of Stomatology,National Clinical Research Center for Oral Diseases,Beijing 100081,China)

机构地区：[1]国家口腔疾病临床医学研究中心,口腔生物材料和数字诊疗装备国家工程研究中心,口腔数字医学北京市重点实验室,北京大学口腔医学院口腔医院修复科,北京100081

出　　处：《生物化学与生物物理进展》2024年第6期1406-1417,共12页Progress In Biochemistry and Biophysics

基　　金：supported by grants from Beijing Natural Science Foundation(7242174,L222030)。

摘　　要：目的本研究旨在明确精氨酸甲基转移酶(PRMT)7在人骨髓间充质干细胞(hBMSCs)成脂分化过程中的变化以及是否调控hBMSCs成脂分化,进而探索相应的调控机制。方法通过定量反转录PCR(qRT-PCR)和蛋白质印迹(Western blot)检测hBMSCs成脂分化过程中PRMT7的变化;通过qRT-PCR和Western blot实验证明PRMT7稳定敲低细胞系构建成功。进行油红O染色和定量分析,以及qRT-PCR和Western blot实验检测PRMT7稳定敲低细胞系成脂分化水平的变化;通过裸鼠体内异位成脂实验,油红O染色检测PRMT7稳定敲低细胞系体内异位成脂的效果;通过qRT-PCR和Western blot证明PRMT7稳定过表达细胞系构建成功。进行油红O染色和定量分析以及qRT-PCR和Western blot实验检测PRMT7稳定过表达细胞系成脂分化水平的变化;通过q RT-PCR和Western blot实验检测敲低PRMT7和过表达PRMT7的细胞中IGF-1表达水平的变化。在PRMT7稳定敲低细胞系中转染siIGF-1并通过qRT-PCR和Western blot检测IGF-1的表达水平验证敲低效率。通过油红O染色和定量分析,qRT-PCR实验检测转染siIGF-1的敲低组hBMSCs成脂分化水平的变化。结果本文发现:在hBMSCs成脂过程中,PRMT7表达水平明显降低(P<0.01);敲低PRMT7后hBMSCs的成脂分化能力增强(P<0.001);敲低PRMT7后hBMSCs的体内异位成脂分化能力增强;过表达PRMT7后hBMSCs的成脂分化能力减弱(P<0.01);PRMT7敲低后IGF-1表达水平增加(P<0.0001);PRMT7过表达后IGF-1表达水平降低(P<0.0001);转染siIGF-1后,各细胞系IGF-1表达水平明显降低(P<0.001);敲低组转染siIGF-1后成脂分化能力明显降低(P<0.01)。结论本研究通过细胞水平和裸鼠皮下移植实验发现PRMT7显著抑制hBMSCs成脂分化,机制研究发现PRMT7对hBMSCs成脂分化的调控作用依赖IGF-1信号通路。上述研究表明,PRMT7可能是治疗相关疾病的潜在分子靶点,为PRMT7和hBMSCs应用于相关疾病治疗提供了新思路。Objective Protein arginine methyltransferases(PRMTs)play pivotal roles in numerous cellular biological processes.However,the precise regulatory effects of PRMTs on the fate determination of mesenchymal stromal/stem cells(MSCs)remain elusive.Our previous studies have shed light on the regulatory role and molecular mechanism of PRMT5 in MSC osteogenic differentiation.This study aims to clarify the role and corresponding regulatory mechanism of PRMT7 during the adipogenic differentiation of bone marrow-derived mesenchymal stem cells(BMSCs).Methods(1)Human bone marrow-derived mesenchymal stem cells(hBMSCs)were cultured in a medium that induces adipogenesis.We used qRT-PCR and Western blot to monitor changes in PRMT7 expression during adipogenic differentiation.(2)We created a cell line with PRMT7 knocked down and assessed changes in PRMT7 expression and adipogenic capacity using Oil Red O staining,qRT-PCR and Western blot.(3)We implanted hBMSCs cell lines mixed with a collagen membrane subcutaneously into nude mice and performed Oil Red O staining to observe ectopic lipogenesis in vivo.(4)A cell line overexpressing PRMT7 was generated,and we examined changes in PRMT7 expression using qRT-PCR and Western blot.We also performed Oil Red O staining and quantitative analysis after inducing the cells in lipogenic medium.Additionally,we assessed changes in PPARγexpression.(5)We investigated changes in insulin-like growth factor 1(IGF-1)expression in both PRMT7 knockdown and overexpressing cell lines using qRT-PCR and Western blot,to understand PRMT7’s regulatory effect on IGF-1 expression.siIGF-1 was transfected into the PRMT7 knockdown cell line to inhibit IGF-1 expression,and knockdown efficiency was confirmed.Then,we induced cells from the control and knockdown groups transfected with siIGF-1 in lipogenic medium and performed Oil Red O staining and quantitative analysis.Finally,we assessed PPARγexpression to explore IGF-1’s involvement in PRMT7’s regulation of adipogenic differentiation in hBMSCs.Results(1)During

关 键 词：精氨酸甲基转移酶7 成脂分化 骨髓间充质干细胞 IGF-1信号通路 

分 类 号：Q254[生物学—细胞生物学] Q28Q291