Title:The Influence of Tetracycline Inducible Targeting Rat PPARγ Gene Silencing on the Osteogenic and Adipogenic Differentiation of Bone Marrow Stromal Cells
Volume: 22
Issue: 41
Author(s): Xiaobo Feng, Xianzhe Liu, Xianyi Cai, Tao Lin, Weihua Xu, Cao Yang, Yongwei Liu, Shuhua Yang and Dehao Fu
Affiliation:
Keywords:
PPARγ, shRNA, Tetracycline, lentiviral vector, Osteogenic differentiation, Adipogenic differentiation, Rat bone marrow stromal
cells.
Abstract: Peroxisome proliferator-activated receptor γ (PPARγ) has been considered as the master regulator for adipogenesis
of bone marrow stromal cells (BMSCs). However, there are few reports regarding the effect of PPARγ gene silencing
on osteogenic and adipogenic differentiation in rat BMSCs, and no reports about tissue targeting and conditional
knockdown of PPARγ gene. In this study, we construct rat PPARγ gene shRNA Tet-on lentiviral vector, the lentiviral
vector facilitated tetracycline (which has the characteristics of bone targeting)-inducible knockdown specific to PPARγ
gene, and transfect it into BMSCs, the silencing effects induced by tetracycline is significant. The expression of the adipogenic
factors adipocyte determination and differentiation-dependent factor 1 (ADD1) and recombinant CCAAT/enhancer
binding protein alpha (C/EBPα) were decreased as measured by RT-PCR and Western blot assay following PPARγ silencing.
In contrast, expression of the osteogenic genes encoding collagen I and Cbfa1/Runx2 were increased. In adipogenic
medium, PPARγ-shRNA transfection reduced the lipid droplet count as measured by Oil red O staining when compared to
the control groups. In osteogenic medium, PPARγ-shRNA increased the activity of alkaline phosphatase and the amount
of calcium deposition as measured by Alizarin red S staining. These results suggest that the rat PPARγ gene shRNA Teton
lentiviral vector decreases adipogenic differentiation and promotes osteogenic differentiation in BMSCs induced by
tetracycline.
