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
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