title: The Influence of Tetracycline Inducible Targeting Rat PPARγ Gene Silencing on the Osteogenic and Adipogenic Differentiation of 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.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Feng
authors.forename: Xianzhe
authors.initials: X
authors.lastname: Liu
authors.forename: Xianyi
authors.initials: X
authors.lastname: Cai
authors.forename: Tao
authors.initials: T
authors.lastname: Lin
authors.forename: Weihua
authors.initials: W
authors.lastname: Xu
authors.forename: Cao
authors.initials: C
authors.lastname: Yang
authors.forename: Yongwei
authors.initials: Y
authors.lastname: Liu
authors.forename: Shuhua
authors.initials: S
authors.lastname: Yang
authors.affiliation: Department of Orthopaedics, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China, Jiefang Avenue, No.1277, 430022, China.
authors.forename: Dehao
authors.initials: D
authors.lastname: Fu
mesh_terms: D050156:Adipogenesis / Q000187:drug effects*; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D005260:Female; D020868:Gene Silencing / Q000187:drug effects*; D057809:HEK293 Cells; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects*; D010012:Osteogenesis / Q000187:drug effects*; D047495:PPAR gamma / Q000037:antagonists & inhibitors* / Q000235:genetics; D012333:RNA, Messenger / Q000037:antagonists & inhibitors / Q000235:genetics; D034741:RNA, Small Interfering / Q000037:antagonists & inhibitors / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D013752:Tetracycline / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D047495:PPAR gamma; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D013752:Tetracycline
keywords: Adipogenic differentiation; Osteogenic differentiation; PPARγ; Rat bone marrow stromal
  cells; Tetracycline; lentiviral vector; shRNA