Abstract
Objectives
To express a TAT-PBX1 fusion protein using a prokaryotic expression system and to explore potential effects of TAT-PBX1 in the proliferation and senescence of human hair follicle-derived mesenchymal stem cells.
Results
The TAT-PBX1 fusion was produced in inclusion bodies and heterogenously expressed in Rosetta (DE3) cells. Immunofluorescence staining showed that TAT-PBX1 fusion proteins were internalized by human hair follicle-derived mesenchymal stem cells. The growth rate of cells was increased after treatment with more than 5.0 μg/mL of TAT-PBX1. The rate of senescence-associated β-galactosidase positive cells was reduced in the 10.0 μg/mL TAT-PBX1 group (28%) than the 0 μg/mL control group (60%). Cells treated with the TAT-PBX1 fusion protein showed higher expression of p-AKT (1.22-fold that of the control), which indicates that TAT-PBX1 activated AKT pathway after cellular uptake.
Conclusions
The TAT-PBX1 fusion protein increased the proliferation of hair follicle mesenchymal stem cells and delayed their senescence by activating the AKT pathway following internalization by cells.
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Acknowledgements
This work was supported by the Jilin Province Science and Technology Development Plan [20190304044YY]; the Innovative special industry fund project in Jilin province [2018C049-2]; the Joint construction project between Jilin province and provincial colleges [SXGJQY2017-12]; and the China Natural National Science Foundation [81573067].
Supplementary information
Supplementary Figure 1—Isolation and characterization of HF-MSCs.
Supplementary Figure 2—Reverse transcriptase-polymerase chain reaction.
Supplementary Materials and Methods 1—Isolation and identification of hHF-MSCs.
Supplementary Materials and Methods 2—Promoter sequence of pNanog-Luc plasmid.
Supplementary Materials and Methods 3—Reverse transcriptase-polymerase chain reaction (RT-PCR).
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Wang, B., Liu, F., Liu, Z. et al. Internalization of the TAT-PBX1 fusion protein significantly enhances the proliferation of human hair follicle-derived mesenchymal stem cells and delays their senescence. Biotechnol Lett 42, 1877–1885 (2020). https://doi.org/10.1007/s10529-020-02909-x
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DOI: https://doi.org/10.1007/s10529-020-02909-x