title: The use of small interfering RNAs to inhibit adipocyte differentiation in human preadipocytes and fetal-femur-derived mesenchymal cells.
abstract: RNA interference (RNAi) has been used in functional genomics and offers innovative approaches in the development of novel therapeutics. Human mesenchymal stem cells offer a unique cell source for tissue engineering/regeneration strategies. The current study examined the potential of small interfering RNAs (siRNA) against human peroxisome proliferator activated receptor gamma (PPARgamma) to suppress adipocyte differentiation (adipogenesis) in human preadipocytes and fetal-femur-derived mesenchymal cells. Adipogenesis was investigated using cellular and biochemical analysis. Transient transfection with PPARgamma-siRNA using a liposomal-based strategy resulted in a significant inhibition of adipogenesis in human preadipocytes and fetal-femur-derived mesenchymal cells, compared to controls (cell, liposomal and negative siRNA). The inhibitory effect of PPARgamma-siRNA was supported by testing human PPARgamma mRNA and adipogenic associated genes using reverse transcription polymerase chain reaction (RT-PCR) to adiponectin receptor 1 and 2 as well as examination of fatty acid binding protein 3 (FABP3) expression, an adipocyte-specific marker. The current studies indicate that PPARgamma-siRNA is a useful tool to study adipogenesis in human cells, with potential applications both therapeutic and in the elucidation of mesenchymal cell differentiation in the modulation of cell differentiation in human mesenchymal cells.
authors.affiliation: Bone and Joint Research Group, Developmental Origins of Health and Disease Division, University of Southampton, Southampton General Hospital, Tremona Road, Southampton SO16 6YD, UK.
authors.forename: Y
authors.initials: Y
authors.lastname: Xu
authors.forename: S-H
authors.initials: SH
authors.lastname: Mirmalek-Sani
authors.forename: X
authors.initials: X
authors.lastname: Yang
authors.forename: J
authors.initials: J
authors.lastname: Zhang
authors.forename: R O C
authors.initials: RO
authors.lastname: Oreffo
mesh_terms: D017667:Adipocytes / Q000166:cytology*; D050156:Adipogenesis; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D048430:Cell Shape; D002478:Cells, Cultured; D000074026:Fatty Acid Binding Protein 3; D050556:Fatty Acid-Binding Proteins / Q000378:metabolism; D005260:Female; D005269:Femur / Q000196:embryology; D005333:Fetus; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology*; D047495:PPAR gamma / Q000235:genetics / Q000378:metabolism; D011247:Pregnancy; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: C493408:FABP3 protein, human; D000074026:Fatty Acid Binding Protein 3; D050556:Fatty Acid-Binding Proteins; D047495:PPAR gamma; D034741:RNA, Small Interfering