Survivin基因在骨髓间充质干细胞中的表达研究

发布时间：2018-04-11 19:40

本文选题：基因 + 骨髓间充质干细胞　；参考：《兰州大学》2009年硕士论文

【摘要】： 目的本研究分离培养骨髓源间充质干细胞(mesenchymal stem cells,MSCs),检测survivin基因在其中的表达情况,并分别通过紫外线照射及粒细胞集落刺激因子刺激后,观察MSCs中survivin基因的表达变化,探讨其意义。方法应用Ficoll密度梯度离心法分离骨髓间充质干细胞,LG-DMEM培养基,15%FBS培养,培养过程中绘制细胞生长曲线,流式细胞术检测其特异性表面标志,并用地塞米松、胰岛素、吲哚美辛诱导成脂,21天后油红O染色照相,采用半定量RT-PCR法检测MSCs中survivin基因在转录水平上的表达情况。并通过紫外线照射、G-CSF刺激等方法处理后,采用半定量RT-PCR法检测其中survivin基因表达的变化。结果原代和传代培养MSCs均保持较强的增殖能力,细胞生长曲线提示第7天传代细胞进入平台期,流式细胞仪检测示CD83、CD45、组织相容性抗原HLA-DR及干细胞表面标志CD34无表达或极低表达,而CD44、CD13、CD71高表达,经地塞米松等诱导后MSCs向脂肪细胞分化,RT-PCR结果显示正常人骨髓间充质干细胞中有survivin基因表达,并且分别经过紫外线照射、G-CSF处理后人骨髓间充质干细胞中survivin基因的表达量有不同程度的提高。结论 Survivin基因不是癌细胞特异性抗凋亡蛋白,在正常人骨髓间充质干细胞中有表达,并且通过紫外线照射、G-CSF刺激等处理后其表达量有不同程度的提高,提示它可能参与了间充质干细胞的生物学行为并影响着间充质干细胞存活。
[Abstract]:PurposeIn this study, mesenchymal stem cells were isolated from bone marrow mesenchymal stem cells and the expression of survivin gene was detected. The changes of survivin gene expression in MSCs were observed after ultraviolet irradiation and granulocyte colony stimulating factor stimulation.MethodBone marrow mesenchymal stem cells were isolated from LG-DMEM medium by Ficoll density gradient centrifugation and cultured in 15s. The cell growth curve was drawn and the specific surface markers were detected by flow cytometry.Indomethacin was induced into lipids for 21 days and oil red O staining was used to detect the expression of survivin gene at the transcriptional level in MSCs by semi-quantitative RT-PCR assay.The changes of survivin gene expression were detected by semi-quantitative RT-PCR after treated by ultraviolet irradiation and G-CSF stimulation.ResultThe cell growth curve indicated that the passage cells entered the plateau phase on the 7th day. Flow cytometry showed that CD83 + CD45, histocompatibility antigen HLA-DR and stem cell surface marker CD34 had no or very low expression.However, CD44, CD13, CD71 were highly expressed. RT-PCR results of MSCs differentiation into adipocytes showed that survivin gene was expressed in normal human bone marrow mesenchymal stem cells.The expression of survivin gene in human bone marrow mesenchymal stem cells was increased by UV irradiation.ConclusionSurvivin gene is not a specific anti-apoptotic protein of cancer cells. It is expressed in normal human bone marrow mesenchymal stem cells.It may be involved in the biological behavior of mesenchymal stem cells and affect the survival of mesenchymal stem cells.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329

【引证文献】