全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

microRNA26b-5p靶向下调p300对人脐带间充质干细胞的增殖作用

, PP. 1614-1618

Keywords: 人脐带间充质干细胞,转录共激活因子p,mirb-p,Nanog

Full-Text   Cite this paper   Add to My Lib

Abstract:

目的探讨潜在microRNA靶向下调p300表达水平对人脐带间充质干细胞(humanumbilicalcordderivedmesenchymalstemcells,hUC-MSCs)增殖的影响。方法通过双荧光素酶报告基因(Firefly/RenillaLuciferase)系统筛选出作用于p3003′UTR的目标microRNA,mimic转染UC-MSCs后RT-qPCR检测miRNA表达水平的变化,Westernblot检测p300蛋白水平变化,RT-qPCR和Westernblot检测多潜能转录因子表达水平的变化,MTT法检测UC-MSCs增殖能力的变化,并比较克隆形成能力的差异。结果证实mir26b-5p通过不完全互补配对结合p300的3′UTR区,显著降低了荧光素酶的相对活性(P<0.05);mir26b-5pmimic转染后抑制p300蛋白表达水平(P<0.05),并下调Nanog基因和蛋白表达水平(P<0.05);mir26b-5pmimic转染后UC-MSCs的增殖受到抑制(P<0.05),克隆形成能力下降(P<0.05)。结论UC-MSCs高表达mir26b-5p能够显著抑制p300蛋白表达和干性相关基因Nanog的表达,并且抑制UC-MSCs的增殖能力和克隆形成能力。

References

[1]  Wang H S, Hung S C, Peng S T, et al. Mesenchymal stem cells in the Wharton’s jelly of the human umbilical cord[J]. Stem Cells, 2004, 22(7): 1330-1337. [2]Melton C, Judson R L, Blelloch R. Opposing microRNA families regulate self-renewal in mouse embryonic stem cells[J]. Nature, 2010, 463(7281): 621-626. [3]Verfaillie C. Adult stem cells: Tissue specific or not?[M]// Lanza R, Atala A. Essentials of Stem Cell Biology. 3rd ed. Oxford: Elsevier Academic Press, 2006: 23-27. [4]Choumerianou D M, Martimianaki G, Stiakaki E, et al. Comparative study of stemness characteristics of mesenchymal cells from bone marrow of children and adults[J]. Cytotherapy, 2010, 12(7): 881-887. [5]Zhong X, Jin Y. Critical roles of coactivator p300 in mouse embryonic stem cell differentiation and Nanog expression[J]. J Biol Chem, 2009, 284(14): 9168-9175. [6]Fang F, Xu Y, Chew K K, et al. Coactivators p300 and CBP maintain the identity of mouse embryonic stem cells by mediating long-range chromatin structure[J]. Stem Cells, 2014, 32(7): 1805-1816. [7]Wang Y, Medvid R, Melton C, et al. DGCR8 is essential for microRNA biogenesis and silencing of embryonic stem cell self-renewal[J]. Nat Genet, 2007, 39(3): 380-385. [8]Riekstina U, Cakstina I, Parfejevs V, et al. Embryonic stem cell marker expression pattern in human mesenchymal stem cells derived from bone marrow, adipose tissue, heart and dermis[J]. Stem Cell Rev, 2009, 5(4): 378-386. [9]Tsai C C, Su P F, Huang Y F, et al. Oct4 and Nanog directly regulate Dnmt1 to maintain self-renewal and undifferentiated state in mesenchymal stem cells[J]. Mol Cell, 2012, 47(2): 169-182. [10]Tsai C C, Chen Y J, Yew T L, et al. Hypoxia inhibits senescence and maintains mesenchymal stem cell properties through down-regulation of E2A-p21 by HIF-TWIST[J]. Blood, 2011, 117(2): 459-469. [11]Greco S J, Liu K, Rameshwar P. Functional similarities among genes regulated by OCT4 in human mesenchymal and embryonic stem cells[J]. Stem Cells, 2007, 25(12): 3143-3154. [12]Chen X, Xu H, Yuan P, et al. Integration of external signaling pathways with the core transcriptional network in embryonic stem cells [J]. Cell, 2008, 133(6): 1106-1117. [13]郭允倩, 陈默, 马天骅, 等. p300通过homeobox结构域增强Nanog的转录激活活性[J]. 中国生物工程杂志, 2009, 29(10): 1-5. ?

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133