Tissue Inhibitor of Matrix Metalloproteinases-1 Knockdown Suppresses the Proliferation of Human Adipose-Derived Stem Cells.
Tissue Inhibitor of Matrix Metalloproteinases-1 Knockdown Suppresses the Proliferation of Human Adipose-Derived Stem Cells.
复制标题
基质金属蛋白酶-1 的组织抑制剂敲低可抑制人脂肪干细胞的增殖
DOI：
10.1155/2016/4761507
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发表时间：
2016
影响因子：
4.3
通讯作者：
Liu H
中科院分区：
文献类型：
--
作者：
Zhang P;Li J;Qi Y;Tang X;Duan J;Liu L;Wu Z;Liang J;Li J;Wang X;Zeng G;Liu H
金属蛋白酶组织抑制剂-1（TIMP-1）是一种多功能的基质金属蛋白酶，参与调节多种细胞的增殖和凋亡。然而，关于TIMP-1表达对脂肪源性干细胞（ADSC）增殖的影响知之甚少。因此，我们首先采用Western blotting检测TIMP-1在ADSCs中的表达，并采用慢病毒介导的shRNA敲除TIMP-1基因。MTT法检测细胞增殖活性，Ki 67染色检测细胞增殖活性。流式细胞术测定细胞周期进程。Western blotting检测TIMP-1基因敲低对p51、p21、细胞周期蛋白E、细胞周期蛋白依赖性激酶2（cyclin-dependent kinase 2，CDK 2）和P-CDK 2表达的影响。结果表明，脂肪干细胞高表达TIMP-1蛋白，TIMP-1基因敲低可抑制脂肪干细胞增殖，并使细胞周期阻滞于G1期，其机制可能与上调p53、p21和P-CDK 2蛋白水平，同时下调cyclin E和CDK 2蛋白水平有关。这些发现表明，TIMP-1作为ADSC中细胞增殖的正调节剂起作用。
Tissue inhibitor of metalloproteinases-1 (TIMP-1) is a multifunctional matrix metalloproteinase, and it is involved in the regulation of cell proliferation and apoptosis in various cell types. However, little is known about the effect of TIMP-1 expression on the proliferation of adipose-derived stem cells (ADSCs). Therefore, TIMP-1 expression in the ADSCs was firstly detected by western blotting, and TIMP-1 gene was knocked down by lentivirus-mediated shRNA. Cell proliferation was then evaluated by MTT assay and Ki67 staining, respectively. Cell cycle progression was determined by flow cytometry. The changes of p51, p21, cyclin E, cyclin-dependent kinase 2 (CDK2), and P-CDK2 caused by TIMP-1 knockdown were detected by western blotting. The results indicated that ADSCs highly expressed TIMP-1 protein, and the knockdown of TIMP-1 inhibited cell proliferation and arrested cell cycle progression at G1 phase in the ADSCs possibly through the upregulation of p53, p21, and P-CDK2 protein levels and concurrent downregulation of cyclin E and CDK2 protein levels. These findings suggest that TIMP-1 works as a positive regulator of cell proliferation in ADSCs.