TAP63γ基因调节Col10a1基因表达及软骨细胞分化成熟  

TAP63γgene regulates Col10a1 gene expression and chondrocyte differentiation and maturation

作　　者：李娜[1] 王倩 夏康 孙晓曦 展丙香 吴璇 陈晨 郑其平 LI Na;WANG Qian;XIA Kang;SUN Xiaoxi;ZHAN Bingxiang;WU Xuan;CHEN Chen;ZHENG Qiping(Department of Blood Transfusion,the First Affiliated Hospital of Anhui Medical University,Hefei,Anhui 230022,China;School of Medicine,Jiangsu University,Jiangsu,Zhenjiang 212013,China)

机构地区：[1]安徽医科大学第一附属医院输血科,安徽合肥230022 [2]江苏大学医学院,江苏镇江212013

出　　处：《国际检验医学杂志》2021年第24期2963-2966,2970,共5页International Journal of Laboratory Medicine

基　　金：国家自然科学基金项目(81702111);江苏省科技计划项目(BE2020679)。

摘　　要：目的拟应用两种软骨细胞模型MCT和ATDC5细胞阐明TAP63γ基因如何调节十型胶原蛋白基因(Col10a1)基因表达及软骨细胞分化、成熟。方法应用荧光定量聚合酶链反应(PCR)技术检测TAP63γ基因和Col10a1基因分别在增殖/肥大状态下小鼠MCT细胞和ATDC5细胞中的表达。应用基因转染,在MCT细胞和ATDC5细胞中过表达或抑制TAP63γ基因的表达,以研究其对Col10a1基因表达的调控作用。在细胞诱导培养的第4、7、14、21天,用阿尔新蓝染色、碱性磷酸酶染色和茜素红染色分别对稳定转染TAP63γ表达质粒的目的组和转染空载体pCMV的对照组及未转染的空白组进行染色,统计分析阳性染色面积,从而确定过表达TAP63γ基因在软骨细胞骨化期间给软骨内成骨所带来的影响。结果与增殖型软骨细胞比较,TAP63γ基因与Col10a1基因在MCT和ATDC5两种肥大型软骨细胞的相对表达量都明显升高(P<0.05)。在MCT细胞和ATDC5细胞中抑制TAP63γ基因的表达,结果导致Col10a1基因的相对表达量明显降低,而过表达TAP63γ基因则明显升高Col10a1基因的相对表达量。阿尔新蓝染色在诱导培养的第7天显示出最强的染色强度,但目的组与对照组没有明显差异;在诱导培养的第21天,碱性磷酸酶染色目的组比对照组显示出更强的染色强度,茜素红染色未见差异。结论TAP63γ基因与两种软骨细胞模型MCT和ATDC5细胞中Col10a1基因的表达一致,TAP63γ基因促进软骨细胞的成熟分化。TAP63γ基因可能与多种转录因子一起构成调节Col10a1基因表达的新型机制进而影响骨骼发育与疾病中软骨细胞成熟的进程。Objective To investigate the in vitro function of TAP63γgene on Col10a1 gene expression and chondrocyte hypertrophic differentiation and maturation using the MCT and ATDC5 chondrocyte cell models.Methods Expression of TAP63γgene and Col10a1 gene in these two proliferative or hypertrophic cells were examined using qRT-PCR.The effects of TAP63γgene on Col10a1 gene expression were evaluated using gene transfection to over-express or inhibit TAP63γgene in MCT and ATDC5 cells.The in vitro effects of TAP63γgene on chondrocyte differentiation was determined by using alcian blue,alkaline phosphatase and alizarin red staining,and analysis the positive staining area of the TAP63γgene expression plasmid cell stably transfected cell(the objective group)and the control cell(the control group)lines after cell induced culture for 4,7,14 and 21 days.Results When compared with their proliferative chondrocyte,TAP63γgene and Col10a1 gene expression in hypertrophic MCT and ATDC5 cells were significantly up-regulated.It leaded to significant down-regulation of Col10a1 gene expression when inhibition of TAP63γgene in MCT and ATDC5 cells,while over-expression of TAP63γgene resulted in significant increase of Col10a1 gene.Alcian blue staining showed the strongest staining intensity in day 7,but the objective group and the control group had no obvious difference.In the induced culture of day 21,objective group showed stronger staining intensity than control group by alkaline phosphatase staining,alizarin red staining did not see the difference.Conclusion TAP63γgene is consistent with Col10a1 gene expression in both chondrocyte models.TAP63γgene promotes chondrocyte hypertrophic differentiation.TAP63γgene may constitute a novel mechanism for regulating Col10a1 gene expression together with several transcription factors,and thus affect bone development and chondrocyte maturation in related skeletal diseases.

关 键 词：Col10a1基因 MCT ATDC5 TAP63γ基因 软骨细胞肥大 

分 类 号：R446.9[医药卫生—诊断学]