AIM: This study is aimed to investigate Runx2’s regulation by phytoestrogens resveratrol and genistein, screen for Runx2 mutation in Chinese cleidocranial dysplasia(CCD) patients, also determine the functional differ...AIM: This study is aimed to investigate Runx2’s regulation by phytoestrogens resveratrol and genistein, screen for Runx2 mutation in Chinese cleidocranial dysplasia(CCD) patients, also determine the functional difference between Runx2’s two major isoforms-type I and type II. METHODS: The simian virus 40 large T antigens (SV40LT) gene was used to establish immortalized hBMSCs and the cell line were then identified by specific surface markers confirmation and pro-longed life span and renewable proliferative capacity illustration. Then we tested resveratrol and genistein’s regulation on the cell line by monitoring several osteoblastic markers and invested the possible involved pathway by western blotting. Subsequently, we determined the functional difference of Runx2’s two major isoforms-type I and type II-in osteoblastic differentiation by RNAi. Also for the first time we screened for the Runx2 mutations in Chinese CCD patients by DNA analysis. RESULTS: We successfully established a permanent hBMSCs by stable transfection of SV40LT. Based on this cell line platform, we illustrated that phytoestrogens resveratrol and genistein regulated Runx2’s expression in osteoblastic differentiation and might partly via MAPK pathway. We also found that type I and type II play a different role in osteoblastic differentiation process, during which type I major increased in the early phase and important for the cell proliferation while type II predominantly functioned in the late stage and necessary for the cell maturation. What’s more, we sequencing coding domains and promoter domains of Runx2 from 9 patients in 3 different CCD familial cases and found two nucleotide changes. CONCLUSION: In this paper, we set up an immortalized hBMSC line by stable transfection of SV40LT which brings convenience for the later studies, reveals that resveratrol and genistein are capable of promote osteoblastic differentiation and are probably via MAPK pathway, also finds that type I major functions in the early stage of osteoblastic differentiation while type II predominantly functions in the late stage, which for the first time distinguishes their effects and gave new hints for bone-related diseases therapies. Moreover, the investigation on RUNX2 mutations in CCD patients adds new records to the repertoire of RUNX2 mutations in China.展开更多
文摘目的了解急性胰腺炎(acute pancreatitis,AP)的生物学和分子发病机制,为预防和治疗AP提供依据。方法从基因表达总库数据库获取基因表达数据集GSE109227、GSE3644和GSE65146,其中包括AP小鼠和对照小鼠胰腺样本的基因表达谱。使用GEO2R鉴定所有差异表达基因(differentially expressed genes,DEGs),并使用基因本体(gene ontology,GO)和京都基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析进行注释。结果在GSE109227、GSE3644和GSE65146数据集中发现38个共同的DEGs,将其构建成1个蛋白质-蛋白质相互作用(protein-proteininteraction,PPI)网络,确定了枢纽蛋白,并提取了功能模块。通过PPI网络分析确定参与粘连接头通路的3个基因(Cdh1、Iqgap1和Actn4),可能为AP的诊断和治疗提供潜在的生物标志物。结论Cdh1、Iqgap1和Actn4基因对AP的发生和发展有影响,这可能为AP的诊断和治疗提供新的靶点。
文摘AIM: This study is aimed to investigate Runx2’s regulation by phytoestrogens resveratrol and genistein, screen for Runx2 mutation in Chinese cleidocranial dysplasia(CCD) patients, also determine the functional difference between Runx2’s two major isoforms-type I and type II. METHODS: The simian virus 40 large T antigens (SV40LT) gene was used to establish immortalized hBMSCs and the cell line were then identified by specific surface markers confirmation and pro-longed life span and renewable proliferative capacity illustration. Then we tested resveratrol and genistein’s regulation on the cell line by monitoring several osteoblastic markers and invested the possible involved pathway by western blotting. Subsequently, we determined the functional difference of Runx2’s two major isoforms-type I and type II-in osteoblastic differentiation by RNAi. Also for the first time we screened for the Runx2 mutations in Chinese CCD patients by DNA analysis. RESULTS: We successfully established a permanent hBMSCs by stable transfection of SV40LT. Based on this cell line platform, we illustrated that phytoestrogens resveratrol and genistein regulated Runx2’s expression in osteoblastic differentiation and might partly via MAPK pathway. We also found that type I and type II play a different role in osteoblastic differentiation process, during which type I major increased in the early phase and important for the cell proliferation while type II predominantly functioned in the late stage and necessary for the cell maturation. What’s more, we sequencing coding domains and promoter domains of Runx2 from 9 patients in 3 different CCD familial cases and found two nucleotide changes. CONCLUSION: In this paper, we set up an immortalized hBMSC line by stable transfection of SV40LT which brings convenience for the later studies, reveals that resveratrol and genistein are capable of promote osteoblastic differentiation and are probably via MAPK pathway, also finds that type I major functions in the early stage of osteoblastic differentiation while type II predominantly functions in the late stage, which for the first time distinguishes their effects and gave new hints for bone-related diseases therapies. Moreover, the investigation on RUNX2 mutations in CCD patients adds new records to the repertoire of RUNX2 mutations in China.
