JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2017, Vol. 55 ›› Issue (7): 23-30.doi: 10.6040/j.issn.1671-7554.0.2016.1682

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Effects of Sirtuin on hypoxia-induced proliferation of human umbilical cord-derived mesenchymal stem cells

WANG Kaimin1, TAN Juanjuan2, YAN Zhiqiang3, LI Zhiqiang1   

  1. 1. Department of Neurosurgery, Shanghai Fengxian District Central Hospital Affiliated to the Third Clinical Medical College of Southern Medical University, Shanghai 201499, China;
    2. School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China;
    3. Department of Central Laboratory, Shanghai Fengxian District Central Hospital Affiliated to the Third Clinical Medical College of Southern Medical University, Shanghai 201499, China
  • Received:2016-12-20 Online:2017-07-10 Published:2017-07-10

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Abstract: Objective To explore the effect of hypoxia on Sirtuin(SIRT)protein expression in human umbilical cord-derived mesenchymal stem cells(hUC-MSCs). Methods hUC-MSCs were isolated and cultured by explant technique. The cells were further expanded and assessed for their morphology and phenotype by flow cytometry. The proliferation of hUC-MSCs under normoxia(21%O2)and hypoxia(5%O2)was evaluated by CCK-8; Cell cycle was analyzed by flow cytometry. Immunofluorescence assay was used to detect the location of SIRT(1-6)under normoxia and hypoxia 山 东 大 学 学 报 (医 学 版)55卷7期 -王凯民,等.Sirtuin在低氧诱导人脐带间充质干细胞增殖中的作用 \=-conditions. Expressions of SIRT(1,2,5)under normoxia and hypoxia conditions for different times(24, 48, 72 h )were detected by Western blotting. Results hUC-MSCs were successfully isolated from umbilical cord. Flow cytometry showed that CD105, CD90, CD44 and CD29 were positive, while CD14, CD19, CD34, CD45 and HLA-DR were negative on hUC-MSCs. The proliferation of hUC-MSCs was enhanced under hypoxia compared with normoxia(all P<0.05). The cells fraction at S phase was also increased under hypoxia compared with normoxia condition after being cultured for 24 h and 48 h(all P <0.05). Immunofluorescence showed that no matter under hypoxia or nomoxia conditions, SIRT1 and SIRT6 were localized in the nucleus, SIRT2 was localized in cytoplasm, and SIRT3, SIRT4, SIRT5 were localized in mitochondria. Western blotting showed the expression of SIRT1 in hUC-MSCs increased under hypoxia after being cultured for 24 h and 48 h(all P<0.05). Similarly, the expression of SIRT5 was increased under hypoxia at all three time points(all P<0.05). The expression of SIRT2 in hypoxia was increased after being cultured for 24 h and 72 h compared with normoxic cells(all P<0.05), however, the expression of SIRT2 under hypoxia condition was lower than that under normoxia condition for 48 h(P<0.05). Conclusion Hypoxia can increase hUC-MSCs’ proliferation. The expressions of SIRT1, SIRT2, SIRT5 were affected by hypoxia, which implies that SIRT1, SIRT2, SIRT5 may be involved in the regulation of hypoxia induced proliferation of hUC-MSCs.

Key words: Human umbilical cord-derived mesenchymal stem cell, Sirtuin, Proliferation, Hypoxia

CLC Number: 

  • R329.2
[1] Han YF, Tao R, Sun TJ, et al. Optimization of human umbilical cord mesenchymal stem cell isolation and culture methods[J]. Cytotechnology, 2013, 65(5): 819-827.
[2] Ding DC, Chang YH, Shyu WC, et al. Human umbilical cord mesenchymal stem cells: a new era for stem cell therapy[J]. Cell Transplant, 2015, 24(3): 339-347.
[3] Kim JH, Park SH, Park SG, et al. The pivotal role of reactive oxygen species generation in the hypoxia-induced stimulation of adipose-derived stem cells[J]. Stem Cells Dev, 2011, 20(10): 1753-1761.
[4] Wagner W, Horn P, Castoldi M, et al. Replicative senescence of mesenchymal stem cells: a continuous and organized process[J]. PloS One, 2008, 3(5): e2213. doi: 10.1371/journal.pone.0002213.
[5] Shibata KR, Aoyama T, Shima Y, et al. Expression of the p16INK4A gene is associated closely with senescence of human mesenchymal stem cells and is potentially silenced by DNA methylation during in vitro expansion[J]. Stem Cells, 2007, 25(9): 2371-2382.
[6] Goossens GH, Bizzarri A, Venteclef N, et al. Increased adipose tissue oxygen tension in obese compared with lean men is accompanied by insulin resistance, impaired adipose tissue capillarization, and inflammation[J]. Circulation, 2011, 124(3): 67-76. doi:10.1161/CIRCULATIONAHA.111.027813.
[7] Marieke RVR, Mensah FKF, Korevaar SS, et al. Effects of hypoxia on the immunomodulatory properties of adipose tissue-derived mesenchymal stem cells[J]. Front Immunol, 2013, 4(1): 203. doi: 10.3389/fimmu.2013.00203.
[8] 陈亭, 周燕, 张治萍,等.间充质干细胞在低氧环境下的增殖、代谢与成骨分化:胎盘羊膜及骨髓来源间充质干细胞的对比[J]. 中国组织工程研究, 2010, 14(6): 957-961. CHEN Ting, ZHOU Yan, ZHANG Zhiping, et al. Proliferation, metabolism, and osteogenic differentiation of mesenchymal stem cells under hypoxia: cell sources of placenta amniotic versus bone marrow[J]. J Clin Rehabil Tis Eng Res, 2010, 14(6): 957-961.
[9] Guarente L. Sirtuins in aging and disease[J]. Cold Spring Harb Symp Quant Biol, 2007, 72(1): 483-488.
[10] Feldman JL, Dittenhafer-Reed KE, Denu JM. Sirtuin catalysis and regulation[J]. J Biol Chem, 2012, 287(51): 42419-42427.
[11] Carafa V, Rotili D, Forgione M, et al. Sirtuin functions and modulation: from chemistry to the clinic[J]. Clin Epigenetics, 2016, 8(1): 1-21.
[12] Kiran S, Anwar T, Kiran M, et al. Sirtuin 7 in cell proliferation, stress and disease: rise of the seventh sirtuin![J]. Cell Signal, 2015, 27(3): 673-682.
[13] 韩潇, 白海, 尹娇娇,等. 氯化钴模拟低氧环境下人脐带间充质干细胞增殖及基因蛋白的表达[J]. 中国组织工程研究, 2015, 19(45): 7268-7273. HAN Xiao, BAI Hai, YIN Jiaojiao, et al. Effect of cobalt chloride-induced hypoxia on proliferation of human umbilical cord-derived mesenchymal stem cells and related gene and protein expressions[J]. J Clin Rehabil Tis Eng Res, 2015, 19(45): 7268-7273.
[14] Tsai CC, Chen YJ, Yew TL, et al. Hypoxia inhibits senescence and maintains mesenchymal stem cell properties through down-regulation of E2A-p21 by HIF-TWIST[J]. Blood, 2010, 117(2): 459-469.
[15] Peng L, Shu X, Lang C, et al. Effects of hypoxia on proliferation of human cord blood-derived mesenchymal stem cells[J]. Cytotechnology, 2016, 68(4): 1-8.
[16] Yuan HF, Zhai C, Yan XL, et al. SIRT1 is required for long-term growth of human mesenchymal stem cells[J]. J Mol Med, 2012, 90(4): 389-400.
[17] Wang X, Ma S, Meng N, et al. Resveratrol exerts dosage-dependent effects on the self-renewal and neural differentiation of hUC-MSCs[J]. Mol Cells, 2016, 39(5): 418-425.
[18] Yuan HF, Zhai C, Yan XL, et al. SIRT1 is required for long[J]. J Mol Med, 2012, 90(4): 389-400.
[19] 徐芬, 严晋华, 梁华,等. SIRT1敲除对C57BL/6J小鼠脂肪细胞分化的影响和机制探讨[J]. 中华医学杂志, 2013, 93(36): 2857-2860. XU Fen, YAN Jinhua, LIANG Hua, et al. SIRT1 knockout impairs the differentiation of adipocyte in C57BL/6J mice and its underlying mechanism[J]. Natl Med J China, 2013, 93(36): 2857-2860.
[20] North BJ, Marshall BL, Borra MT,et al. The human Sir2 ortholog, SIRT2, is an NAD+-dependent tubulin deacetylase[J]. Mol Cell, 2003, 11(2): 437-444.
[21] Peck B, Chen CY, Ho KK, et al. SIRT inhibitors induce cell death and p53 acetylation through targeting both SIRT1 and SIRT2[J]. Mol Cancer Ther, 2010, 9(4): 844-855.
[22] 马珊珊, 崔渊博, 姚宁,等. sirt2基因高表达对人脐带间充质干细胞衰老的抑制作用[J]. 郑州大学学报(医学版), 2016, 51(1): 10-14. MA Shanshan, CUI Yuanbo, YAO Ning, et al. Inhibition of sirt2 overexpression on aging of human umbilical cord mesenchymal stem cells[J]. Journal of Zhengzhou University(Medical Sicences), 2016, 51(1): 10-14.
[23] Si X, Chen W, Guo X, et al. Activation of GSK3β by sirt2 is required for early lineage commitment of mouse embryonic stem cell[J]. PloS One, 2013, 8(10): e76699-e76699. doi: 10.1371/journal.pone.0076699.
[24] Jing E, Gesta S, Kahn CR. SIRT2 regulates adipocyte differentiation through FoxO1 acetylation/deacetylation[J]. Cell Metab, 2007, 6(2): 105-114.
[25] Seo KS, Park JH, Heo JY, et al. SIRT2 regulates tumour hypoxia response by promoting HIF-1α hydroxylation[J]. Oncogene, 2015, 34(11): 1354-1362.
[26] Lin ZF, Xu HB, Wang JY, et al. SIRT5 desuccinylates and activates SOD1 to eliminate ROS[J]. Biochem Biophys Res Commun, 2013, 441(1): 191-195.
[27] Hsu YC, Wu YT, Yu TH, et al. Mitochondria in mesenchymal stem cell biology and cell therapy: from cellular differentiation to mitochondrial transfer[J]. Semin Cell Dev Biol, 2016, 52: 119-131. doi: 10.1016/j.semcdb.2016.02.011.
[28] Schlicker C, Gertz M, Papatheodorou P, et al. Substrates and regulation mechanisms for the human mitochondrial sirtuins Sirt3 and Sirt5[J]. J Mol Biol, 2008, 382(3): 790-801.
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