Changes in bone marrow mesenchymal stem cells osteogenesis by the regulation of Lnk/stem cell factor-cKit signaling.
- Author:
Hao LI
;
Hailun ZHOU
;
Qi WANG
;
Wei LI
- Publication Type:Journal Article
- MeSH: Alkaline Phosphatase; Animals; Bone Marrow Cells; Cell Differentiation; Cells, Cultured; Diabetes Mellitus, Experimental; Hematopoietic Stem Cells; Mesenchymal Stromal Cells; Osteocalcin; Osteogenesis; RNA Interference; Rats; Signal Transduction; Stem Cell Factor; Transfection
- From: West China Journal of Stomatology 2015;33(6):633-637
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEChanges in the osteogenesis of diabetic rat bone marrow mesenchymal stem cells (BMSCs) by the regulation of Lnk/stem cell factor (SCF)-cKit signaling were investigated.
METHODSBMSCs were isolated from diabetic rats and identified by immunocytochemical staining. These cells were divided into the control group (untransfected group), negative control group (transfected with control plasmid), and RNA interference group (transfected with Lnk-targeting RNA interference plasmid). Western blot was performed to analyze the effect of interference. The BMSCs were induced for osteogenic differentiation under diabetic conditions, and Western blot was used to examine the expressions of Lnf, SCF, and cKit in Lnk/SCF-cKit signaling and osteogenic proteins alkaline phosphatase (ALP), osteocalcin (OCN), and collagen type I al (ColIal).
RESULTSIsolated cells expressed CD₄₄ and CD₉₀ but not CD₁₁b or CD₄₅. This phenomenon was characteristic of BMSCs. Compared with other diabetic BMSCs, cells in the RNA interference group expressed low Lnk but high SCF and cKit (P < 0.05). Thereafter, 28 days after induction of osteogenic differentiation, cells in the RNA interference group expressed low Lnk but high SCF, cKit, ALP, OCN, and ColIal compared with other diabetic BMSCs (P < 0.05).
CONCLUSIONThe inhibition of Lnk expression and activation of SCF-cKit pathway may improve the osteogenic differentiation of BMSCs under diabetic conditions.