沉默信息调节因子1基因敲减后ATDC5小鼠软骨细胞中被激活细胞退变相关信号通路及差异因子的表达  

Activation of chondrocyte degeneration related signaling pathways and expression of differential factors in ATDC5 mouse cells after silent information regulator 1 knockdown

作　　者：解笑宸[1,2] 杨琪 翁鉴 曾晖[1,2] 康斌 刘佩[1,2] 钟华戈 于斐[1,2] Xie Xiaochen;Yang Qi;Weng Jian;Zeng Hui;Kang Bin;Liu Pei;Zhong Huage;Yu Fei(Department of Bone&Joint Surgery,Peking University Shenzhen Hospital,Shenzhen 518036,Guangdong Province,China;National&Local Joint Engineering Research Center of Orthopaedic Biomaterials,Shenzhen 518036,Guangdong Province,China;Department of Medical Ultrasound,Peking University Shenzhen Hospital,Shenzhen 518036,Guangdong Province,China;Department of Gastrointestinal Surgery,Guangxi Medical University Affiliated Tumor Hospital,Nanning 530021,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]北京大学深圳医院骨关节科,广东省深圳市518036 [2]骨科生物材料国家地方联合工程研究中心,广东省深圳市518036 [3]北京大学深圳医院超声诊断科,广东省深圳市518036 [4]广西医科大学附属肿瘤医院胃肠外科,广西壮族自治区南宁市530021

出　　处：《中国组织工程研究》2022年第32期5112-5118,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(82102568);国家自然科学基金(82102076),项目负责人:杨琪;国家自然科学基金(82172432);国家自然科学基金(82001319),项目负责人:翁鉴;广东省基础与应用基础研究基金(2021A1515012586,2019A1515110983);白求恩·石药骨质疏松科研基金项目(G-X-2020-1107-21),项目负责人:于斐;广东省基础与应用基础研究基金(2019A1515011290),项目负责人:曾晖。

摘　　要：背景:软骨细胞退变常导致骨关节炎、椎间盘退变、半月板退变等骨科常见疾病,沉默信息调节因子1(silent information regulator 1,SIRT1)基因与软骨细胞退变关系密切,当敲减ATDC5小鼠软骨细胞系中SIRT1基因后,可通过软骨细胞退变相关信号通路的作用影响细胞功能,进而影响骨科疾病的进程。目的:探讨SIRT1基因敲减后ATDC5小鼠软骨细胞中显著激活的软骨细胞退变相关信号通路情况及差异因子的表达情况。方法:携载或不携载SIRT1基因的慢病毒转染ATDC5细胞后,分为对照组(转染携载阴性对照慢病毒ATDC5细胞组)和实验组(转染携载SIRT1基因慢病毒ATDC5细胞组)。提取不同组别细胞中总RNA并进行质检,基因芯片检测细胞中编码RNA核酸的表达情况,结合生物信息学分析,寻找显著激活的软骨细胞退变相关信号通路,并对其中部分信号通路的差异变化因子进行阐述。结果与结论:①发现SIRT1基因敲减后ATDC5细胞中被显著激活的信号通路有42条;②选择了其中被显著激活的5条与软骨细胞退变相关且研究较少的信号通路:肝细胞生长因子信号通路(ETS1、PIK3CA、NRAS、PIK3C2A、FGFR2、ELF1、FGFR3、CDKN1A、AKT3、FRS2、PRKD3、MAP3K2)(Ratio=0.1390,Z-score=2.673)、神经调节因子信号通路(RPS6KB1、STAT5A、MTOR、BTC、HBEGF、RNF41)(Ratio=0.1360,Z-score=2.309)、胰岛素受体信号通路(TSC1、EIF4EBP1、PRKAR2B、PPP1R12A、TSC2)(Ratio=0.1060,Z-score=2.138)、趋化因子受体4信号通路(PAK4、EGR1、RHOJ、RHOB、PLCB1、GNG5)(Ratio=0.0970,Z-score=2.500)、肌动蛋白细胞骨架信号通路(ABI2、BRK1、PIP4K2B、MYLK、IQGAP1、ARPC1A、ACTA2、EZR、SSH2、ACTG1、IQGAP3)(Ratio=0.0921,Z-score=2.236)进行汇报,从而为研究软骨细胞退变相关疾病提供新颖靶点。BACKGROUND:Chondrocytes degeneration often leads to common orthopedic diseases such as osteoarthritis,inte rvertebral disc degeneration and meniscus degeneration.SIRT1 gene is closely related to chondrocytes degeneration.When SIRT1 gene is knocked down in ATDC5 mouse chondrocytes,it can affect cell function via signaling pathways related to chondrocyte degeneration,and then affect the process of orthopedic diseases.OBJECTIVE:To investigate the significantly activated signaling pathways related to chondrocyte degene ration and the expression of differential factors in ATDC5 mouse chondrocytes after SIRT1 gene knockdown.METHODS:The lentivirus carrying SIRT1 gene or not was transfected into ATDC cells.The transfected cells were divided into control(transfected with negative control lentivirus) and expe riment group(transfected with lentivirus carrying SIRT1 gene).Total RNA was extra cted for quality testing.Gene chip was used to detect the expression of coding RNA in the cells.Combined with bioinformatics analysis,we identified significantly activated signaling pathways related to chondrocyte degeneration,and elaborated differentially expressed factors in some signaling pathways.RESULTS AND CONCLUSION:Fo rty-two signaling pathways we re significantly activated in ATDC5 cells after SIRT1 gene knockdown.Among them,we selected five sign aling pathways related to chondrocyte degeneration and repo rted less,including hepatocyte growth fa ctor signaling pathway(ETS1,PIK3 CA,NRAS,PIK3C2A,FGFR2,ELF1,FGFR3,CDKN1A,AKT3,FRS2,PRKD3,MAP3K2;Ratio=0.139 0,Z-score=2.673),neuregulin signaling pathway(RPS6 KB1,STAT5 A,MTO R,BTC,HBEG F,RNF41;Ratio=0.136 0,Z-score=2.309),insulin receptor signaling pathway(TSC1,EIF4EBP1,PRKAR2 B,PPP1R12A,TSC2;Ratio=0.106 0,Z-score=2.138),chemokine receptor 4 signaling pathway(PAK4,EGR1,RHOJ,RHOB,PLCB1,GNG5;Ratio=0.097 0,Z-score=2.500),actin cytos keleton signaling pathway(ABI2,BRK1,PIP4 K2B,MYLK,IQGAP1,ARPC1 A,ACTA2,EZR,SSH2,ACTG1,IQGAP3;Ratio=0.092 1,Z-score=2.236),so as to provide novel targets for tr

关 键 词：SIRT1 ATDC5细胞 软骨细胞退变 基因芯片 细胞实验 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]