Cai Jun,Qu Jifu.Construction of SLPI gene siRNA eukaryotic expression vector and its role in modulating proliferation of dermal multipotent stem cells in rats[J].J Amry Med Univ (J Third Mil Med Univ),2016,38(04):356-360.
Cai Jun,Qu Jifu.Construction of SLPI gene siRNA eukaryotic expression vector and its role in modulating proliferation of dermal multipotent stem cells in rats[J].J Amry Med Univ (J Third Mil Med Univ),2016,38(04):356-360.DOI:
Objective To determine the effect of secretory leukocyte protease inhibitor (SLPI) on promoting proliferation of dermal multipotent stem cells (DMSCs). Methods DMSCs were isolated from neonatal rats by enzyme digestion
and then purified and amplified for further research. Based on successful construction of SLPI o
verexpression vector
SLPI gene siRNA expression vector was picked out according to RT-PCR and Western blot identification. DMSCs were transformed with recombinant expression plasmids of SLPI gene
and the effect of SPLI gene on promoting the proliferation of DMSCs was observed using CCK-8 assay and flow cytometry. Results DMSCs were successfully isolated from the dermis. The eukaryotic expression vector and siRNA expression vector of SLPI gene were constructed. The transformed DMSCs were divided into 3 groups: overexpression group
interference group
and control group (transformation with blank vector). The results of CCK-8 assay showed that
in 48
72 h and 96 h after transformation
the proliferation of DMSCs in the overexpression group was faster than that in the interference group (P0.05)
and in 72 and 96 h after transformation
DMSCs in the overexpression group were significantly different from those in the control group (P0.05)
which suggested that SLPI gene might promote the proliferation of DMSCs. The results of flow cytometry showed that the cells in the G
1
phase in the overexpression group were significantly different from those in the interference group (P0.05)
meanwhile
the cells in the S phase and G
2
phase were significantly different from those in the control group (P0.05). Conclusion SLPI gene plays a role in promoting the proliferation of DMSCs
and promotes cell cycle transformation from G
1
phase to S phase and G
2
phase
which suggests that SLPI may promote proliferation of DMSCs via regulating cell cycle.