Effect of p53 inhibitor on viability of human bone marrow mesenchymal stem cells in late-phase amplification

doi:10.3969/j.issn.2095-4344.2015.23.002

Abstract

Abstract:

BACKGROUND: It is not fully understood that whether p53 inhibitor can directly intervene in the viability of bone marrow mesenchymal stem cells and the possible mechanism.
OBJECTIVE: To investigate the effect of p53 inhibitor, PFT-α, on the aging process of bone marrow mesenchymal stem cells in late-phase amplification and to discover the key target to delay the replicative senescence of human bone marrow mesenchymal stem cells.
METHODS: The expression levels of p53, p21, and p15 mRNA in human bone marrow mesenchymal stem cells in both early and late-phase amplification were detected by quantitative PCR assay. Then, human bone marrow mesenchymal stem cells in late-phase amplification were respectively treated with 20 µmol/L PFT-α or an equivalent amount of dimethyl sulfoxide for 2 weeks. The positive rate of aging cells was determined by SA-β-Gal staining. The apoptosis was detected by TUNEL staining. Human bone marrow mesenchymal stem cells were treated with 300 µmol/L H2O2 for 30 minutes, and then cellular anti-oxidative stress capacity was detected by cell counting kit-8 assay.
RESULTS AND CONCLUSION: The quantitative PCR assay showed that the mRNA expression level of p15,
p21 and p53 in human bone marrow mesenchymal stem cells in late-phase amplification was significantly increased (1.45±0.23), (1.51±0.14) and (1.78±0.14) times as much as that in early phase amplification (P < 0.05). The positive rate of aging cells in PFT-α group was significantly lower than that in the dimethyl sulfoxide group [(41±5)% vs. (63±7)%, P < 0.05)]. However, there was no significant difference in apoptosis rate between PFT-α group and dimethyl sulfoxide group. After treatment with H₂O₂, the absorbance value in the PFT-α group was (1.27±0.13) times as much as that in the dimethyl sulfoxide group (P < 0.001). The above results demonstrate that the activation of p53 signaling pathway may be an important factor of causing aging of human bone marrow mesenchymal stem cells. Application of p53 inhibitor PFT-α can enhance the anti-oxidative stress capacity of human bone marrow mesenchymal stem cells in late phrase amplification.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: 干细胞, 骨髓干细胞, 复制性衰老, 骨髓间充质干细胞, p53, PFT-α, 细胞凋亡, 国家自然科学基金

CLC Number:

R394.2

He Ze-bin, Zhao Yun-he, Yang Gui-jiao, Lu Li. Effect of p53 inhibitor on viability of human bone marrow mesenchymal stem cells in late-phase amplification [J]. Chinese Journal of Tissue Engineering Research, 2015, 19(23): 3616-3620.

Figures/Tables 4

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