PMID- 18197755
OWN - NLM
STAT- MEDLINE
DCOM- 20080421
LR  - 20220227
IS  - 1523-4681 (Electronic)
IS  - 0884-0431 (Linking)
VI  - 23
IP  - 2
DP  - 2008 Feb
TI  - Osteogenic differentiation of human adipose tissue-derived stem cells is 
      modulated by the miR-26a targeting of the SMAD1 transcription factor.
PG  - 287-95
LID - 10.1359/jbmr.071011 [doi]
AB  - The molecular mechanisms that regulate hADSC differentiation toward osteogenic 
      precursors and subsequent bone-forming osteoblasts is unknown. Using osteoblast 
      precursors obtained from subcutaneous human adipose tissue, we observed that 
      microRNA-26a modulated late osteoblasts differentiation by targeting the SMAD1 
      transcription factor. INTRODUCTION: Elucidation of the molecular mechanisms 
      guiding human adipose tissue-derived stem cells (hADSCs) differentiation is of 
      extreme importance for improving the treatment of bone-related diseases such as 
      osteoporosis. The aim of this study was to identify microRNA as a regulator of 
      the osteogenic differentiation of hADSCs. MATERIALS AND METHODS: Osteoblast 
      differentiation of hADSCs was induced by treatment with dexamethasone, ascorbic 
      acid, and beta-glycerol phosphate. The expression of osteoblastic phenotype was 
      evaluated after the induction by simultaneous monitoring of alkaline phosphatase 
      activity, the expression of genes involved in osteoblastic differentiation by 
      real-time RT-PCR, and mineralization at the same time. MicroRNA expression was 
      determined by Northern blot, and transfection of both antisense miR-RNA and 
      sensor plasmids was done to validate the inhibitory role of microRNA during hADSC 
      osteogenesis. Western blot was used to determine the expression levels of the 
      SMAD1 protein. qRT-PCR analysis was used to compare the expression patterns of 
      osteoblastic markers in transfected cells. RESULTS AND CONCLUSIONS: We analyzed 
      the role of microRNA 26a (miR-26a) during differentiation of hADSCs. Northern 
      blot analysis of miR-26a during hADSC differentiation showed increased 
      expression, whereas expression of the SMAD1 protein was complementary to that of 
      miR-26a. Because the highest expression of miR-26a and the lowest expression of 
      SMAD1 protein were reached at hADSC terminal differentiation, we carried out our 
      study during the late stages of hADSC differentiation. The inhibition of miR-26a, 
      by 2'-O-methyl-antisense RNA, increased protein levels of its predicted target, 
      SMAD1 transcription factor, in treated osteoblasts, upregulating bone marker 
      genes and thus enhancing osteoblast differentiation. Our data suggest a role for 
      miR-26a in the differentiation induced by treatment with dexamethasone, ascorbic 
      acid, and beta-glycerol phosphate of hADSCs toward the osteogenic lineage by 
      targeting its predicted target, the SMAD1 protein. This study contributes to a 
      better knowledge of molecular mechanisms governing hADSC differentiation by 
      proposing a microRNA-based control of late differentiation.
FAU - Luzi, Ettore
AU  - Luzi E
AD  - Metabolic Bone Unit, Laboratory of Molecular Genetics, Department of Internal 
      Medicine, University of Florence, Florence, Italy.
FAU - Marini, Francesca
AU  - Marini F
FAU - Sala, Silvia Carbonell
AU  - Sala SC
FAU - Tognarini, Isabella
AU  - Tognarini I
FAU - Galli, Gianna
AU  - Galli G
FAU - Brandi, Maria Luisa
AU  - Brandi ML
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - J Bone Miner Res
JT  - Journal of bone and mineral research : the official journal of the American 
      Society for Bone and Mineral Research
JID - 8610640
RN  - 0 (3' Untranslated Regions)
RN  - 0 (MIRN26A microRNA, human)
RN  - 0 (MicroRNAs)
RN  - 0 (RNA, Messenger)
RN  - 0 (Smad1 Protein)
SB  - IM
MH  - *3' Untranslated Regions/metabolism
MH  - Adipose Tissue/*cytology
MH  - Cell Differentiation/physiology
MH  - Cells, Cultured
MH  - Humans
MH  - MicroRNAs/genetics/*metabolism
MH  - Osteoblasts/cytology/*metabolism
MH  - RNA, Messenger/biosynthesis
MH  - Smad1 Protein/biosynthesis/*metabolism
MH  - Stem Cells/cytology/*metabolism
EDAT- 2008/01/17 09:00
MHDA- 2008/04/22 09:00
CRDT- 2008/01/17 09:00
PHST- 2008/01/17 09:00 [pubmed]
PHST- 2008/04/22 09:00 [medline]
PHST- 2008/01/17 09:00 [entrez]
AID - 10.1359/jbmr.071011 [doi]
PST - ppublish
SO  - J Bone Miner Res. 2008 Feb;23(2):287-95. doi: 10.1359/jbmr.071011.