TGF-β1对hUC-MSC增殖、细胞外基质表达和基因表达的影响  

Effect of TGF-β1 on Proliferation and Extracellular Matrix and Gene Expressions of hUC-MSC

作　　者：赵洁[1] 李栋[2] 时庆[2] 汪大琨[2] 鞠秀丽[1] 

机构地区：[1]山东大学齐鲁医院儿科,山东济南250012 [2]山东大学齐鲁医院低温医学研究室,山东济南250012

出　　处：《中国实验血液学杂志》2011年第6期1466-1471,共6页Journal of Experimental Hematology

摘　　要：本研究探索转化生长因子-β1(TGF-β1)对人脐带间充质干细胞(hUC-MSC)增殖、细胞外基质(ECM)表达和基因表达的影响。采用植块法从人脐带中分离、培养hUC-MSC并进行表型鉴定,用WST法检测不同浓度的TGF-β1对hUC-MSC增殖的影响,利用qRT-PCR和免疫组织化学方法检测不同浓度的TGF-β1对hUC-MSC细胞外基质表达的影响,利用表达基因芯片技术检测hUC-MSC在TGF-β1作用下基因表达谱的变化。结果表明,细胞增殖实验中0.1-20.0 ng/ml TGF-β1浓度条件下培养24、48和72小时,hUC-MSC增殖能力变化均不显著,但在0.1 ng/ml组均会出现相对峰值。0.5 ng/ml的TGF-β1促进Col-Ⅰ、MMP-2、MMP-9和TIMP-2的mRNA表达达到峰值;而0.1 ng/ml的TGF-β1促进其它成分:Col-Ⅳ、FN、LN、Integrin、Tenascin-C、MMP-1和TIMP-1的mRNA表达达到峰值。0.5和0.1 ng/ml的TGF-β1分别促进Col-Ⅰ和Col-Ⅳ少量表达,均促进FN强烈表达,但均不能使LN明显表达。基因芯片结果显示,在上调基因中有9个基因与细胞运动、迁移相关;Pathway和Go的统计分析显示,差异基因涉及转录、翻译、生物合成、代谢、信号转导、细胞迁移、细胞间连接等多方面。结论:0.1 ng/ml的TGF-β1能较好地促进hUC-MSC的生长。不同浓度的TGF-β1能分别促进ECM不同成分表达,TGF-β1对hUC-MSC的转录、翻译、生物合成、代谢、信号转导、细胞迁移、细胞间连接等多方面在基因水平上具有调节作用。This study was purposed to investigate the effects of transforming growth factor-β1（TGF-β1） on proliferation and extracellular matrix（ECM） and gene expressions of human umbilical cord derived mesenchymal stem cells（hUC-MSC）.The hUC-MSCs were isolated and cultured by transplantation technique from umbilical cords.Surface antigens of the fifth passage cultured hUC-MSCs were detected by flow cytometry.The effects of TGF-β1 0.1-20.0 ng/ml on hUC-MSC proliferation were determined by Cell Counting Kit-8.The mRNA expression of ECM components and the components involved in the mechanism of ECM production regulation were detected by qRT-PCR.Expressions of collagen Ⅰ（Col-Ⅰ）,collagen Ⅳ（Col-Ⅳ）,fibronectin（FN） and laminin（LN） were observed by immunocytochemistry.The gene expression profiles of hUC-MSC in response to TGF-β1 were examined by microarray.The results showed that no significant changes of the OD values were observed with the increasing doses of TGF-β1,while all OD values of 0.1 ng/ml group showed relative peaks at 24,48 and 72 hour time points.The mRNA expression level of Col-Ⅰ,MMP-2,MMP-9 and TIMP-2 increased to maximum in 0.5 ng/ml group,and the mRNA expression of Col-Ⅳ,FN,LN,Integrin,Tenascin-C,MMP-1 and TIMP-1 reached the peak level in 0.1 ng/ml group.Immunocytochemical analysis for Col-Ⅰ and Col-Ⅳ revealed that their production increased slightly in 0.5 and 0.1 ng/ml groups,respectively,while immunostaining for FN showed diffuse cell membrane and cytoplasmic staining randomly distributed,but LN staining was absent in both groups.Microarray analysis showed that 9 genes closely related to cell movement and migration were up-regulated.Analysis by Pathway and Go indicated that differentially expressed genes were involved in transcription,translation,biosynthesis,metabolism,signal transduction,migration and adhering junction etc.It is concluded that TGF-β1 0.1 ng/ml promotes the proliferation of hUC-MSC.TGF-β1 of different concentrations upregulates the expression

关 键 词：脐带间充质干细胞 转化生长因子-Β1 细胞外基质 基因表达谱 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]