软骨细胞中SIRT1基因敲减后作用状态不明确信号通路及对相关疾病或功能的影响  

Exploration of signaling pathways with unclear action status and possible effects on related diseases or functions after knockdown of silencing information regulator 1 gene in chondrocytes

作　　者：叶海明[1,2,3] 曾晖 杨琪[4] 张耕 翁鉴 于斐 Ye Haiming;Zeng Hui;Yang Qi;Zhang Geng;Weng Jian;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;Shenzhen Key Laboratory of Orthopaedic Diseases and Biomaterials Research,Shenzhen 518036,Guangdong Province,China;Department of Medical Ultrasound,Peking University Shenzhen Hospital,Shenzhen 518036,Guangdong Province,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]深圳市骨科疾病与生物材料研究重点实验室,广东省深圳市518036 [4]北京大学深圳医院超声影像科,广东省深圳市518036

出　　处：《中国组织工程研究》2024年第20期3123-3129,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(82102568),项目负责人:于斐;广东省基础与应用基础研究基金(2021A1515012586,2022A1515220111),项目负责人:于斐;北京大学深圳医院科研启动金项目(KYQD2021099),项目负责人:于斐;深圳“医疗卫生三名工程”项目(SZSM201612092),项目负责人:曾晖;深圳市医学重点学科建设经费(SZXK023),项目负责人:曾晖;深圳市高水平医院建设专项经费,项目负责人:曾晖。

摘　　要：背景:沉默信息调节因子1以去乙酰化的方式调控软骨细胞中相关蛋白的功能,参与软骨细胞增殖分化,促进软骨缺损修复。目的:利用高通量技术筛选软骨细胞中沉默信息调节因子1基因敲减后作用状态不明确的信号通路以及产生变化的疾病或功能。方法:取处于对数生长期的小鼠ATDC5软骨细胞,分2组转染:对照组转染沉默信息调节因子1基因敲减阴性对照慢病毒,实验组转染沉默信息调节因子1基因敲减慢病毒。转染72 h后,利用小鼠基因表达谱芯片GeneChip®Mouse Genome 4302.0 Array检测mRNA的基因层面变化情况,应用生物信息学技术筛选激活或抑制不明确的信号通路以及这些信号通路相关因子,并分析疾病或功能模块的富集情况。结果与结论:①沉默信息调节因子1基因敲减后,小鼠ATDC5软骨细胞中激活或抑制状态不明确的信号通路有245条;②根据-log(P-value)排序选择排前20的激活或抑制状态不明确信号通路中的因子情况进行报道,包括IGFBP4、TGFBR1、CTGF、COL4A5、LHX2、IL1RL1、KLF6等;③根据-log(P-value)进行排序,细胞生长和增殖、生物体存活和细胞死亡与存活等14个疾病和功能密切相关模块明显变化;根据差异基因数量排序,生物体损伤和异常、癌症和细胞死亡与存活3个疾病和功能密切相关模块明显变化;根据-log(P-value)与差异基因数量综合排序,固有免疫应答这一疾病和功能模块被显著激活。BACKGROUND:silencing information regulatory 1(SIRT1)regulates the function of related proteins in chondrocytes in a deacetylated manner and participates in chondrocyte proliferation and differentiation,thereby promoting cartilage defect repair.OBJECTIVE:To screen for signaling pathways with unclear action status after SIRT1 gene knockdown in chondrocytes,as well as diseases or functions that produce changes using high-throughput technology.METHODS:ATDC5 chondrocytes from mice in logarithmic growth phase were divided into two groups:the cells were transfected with SIRT1 gene knockdown negative control lentivirus in control group and SIRT1 gene knockdown lentivirus in experimental group.GeneChip®Mouse Genome 4302.0 Array was used to detect the mRNA expression at 72 hours after transfection.Applied bioinformatics technology was also used to screen for unclear activation or inhibition signaling pathways and their related factors.Moreover,enrichment of disease or function modules was analyzed.RESULTS AND CONCLUSION:After knocking down the SIRT1 gene,there were 245 signaling pathways with unclear activation or inhibition status in the mouse ATDC5 chondrocytes.According to the ranking of-Log(P-value),we reported the factors in the top 20 signaling pathways with unclear activation or inhibition status,including IGFBP4,TGFBR1,CTGF,COL4A5,LHX2,IL1RL1,and KLF6.According to the ranking of-Log(P-value),there were significant changes in 14 disease or function modules,including cellular growth and proliferation,organism survival,cell death and survival.According to the number of differentially expressed genes,there were significant changes in three disease or function modules,including organismal injury and abnormalities,cancer,and cell death and survival.According to the comprehensive ranking of-Log(P-value)and the number of differentially expressed genes,the disease or function module related to intrinsic immune response was significantly activated.

关 键 词：高通量测序 沉默信息调节因子1 软骨缺损 信号通路 疾病或功能 

分 类 号：R459.9[医药卫生—治疗学] R319[医药卫生—临床医学] R684.3