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PMID: 21794839 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Drosha regulates hMSCs cell cycle progression through a miRNA independent mechanism.

The international journal of biochemistry & cell biology ·Vol. 43 ·No. 11 ·2011-11-00 ·Pages 1563-72

Oskowitz AZ, Penfornis P, Tucker A, Prockop DJ, Pochampally R

Abstract

Recently we demonstrated that the miRNA regulate human mesenchymal stem cells (hMSCs) differentiation. To determine the role of the miRNA pathway in hMSCs proliferation, Drosha and Dicer knockdown hMSCs were generated using a lentiviral based tetracycline inducible shRNA. hMSCs with reduced Drosha expression had a significantly reduced proliferation rate, while hMSCs with reduced Dicer expression displayed a proliferation rate similar to untransduced cells. Cell cycle analysis identified that unlike Dicer knockdown, Drosha knockdown hMSCs contained an increased number of G1 phase cells, with a reduced level of cells in S phase, compared to controls. ELISAs of hMSCs revealed decreased levels of pRB and stable levels of total RB with Drosha knockdown. Two key regulators of the G1/S phase transition, cyclin dependent kinase inhibitor 2A (p16) and cyclin dependent kinase inhibitor 2B (p15), were increased in Drosha knockdown cells but not in Dicer knockdown. Transcripts of 28S and 18S rRNA were significantly reduced in Drosha knockdown hMSCs, with no change in rRNA levels in Dicer knockdown hMSCs. 45S pre-rRNA transcripts were not significantly different in either knockdown model. The above results indicate that Drosha modifies hMSCs proliferation through a miRNA independent mechanism, potentially by regulating rRNA processing.

MeSH Terms
Cell Cycle/genetics Cell Proliferation Cells, Cultured Cyclin-Dependent Kinase Inhibitor p15/genetics,metabolism Cyclin-Dependent Kinase Inhibitor p16/genetics,metabolism DEAD-box RNA Helicases/deficiency,genetics Gene Silencing Genetic Vectors Humans Lentivirus Mesenchymal Stem Cells/cytology,metabolism MicroRNAs/genetics,metabolism RNA, Messenger/genetics RNA, Ribosomal/genetics,metabolism RNA, Small Interfering/genetics Reverse Transcriptase Polymerase Chain Reaction Ribonuclease III/deficiency,genetics Transduction, Genetic Up-Regulation
Chemicals
Cyclin-Dependent Kinase Inhibitor p15 Cyclin-Dependent Kinase Inhibitor p16 MicroRNAs RNA, Messenger RNA, Ribosomal RNA, Small Interfering DICER1 protein, human DROSHA protein, human Ribonuclease III DEAD-box RNA Helicases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Oskowitz Adam Z
Center for Stem Cell Research and Regenerative Medicine, Tulane University Health Sciences Center, New Orleans, LA 70112, USA.
Penfornis Patrice
Tucker Alan
Prockop Darwin J
Pochampally Radhika
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Article Info
Journal
The international journal of biochemistry & cell biology
Abbr.
Int J Biochem Cell Biol
ISSN
1878-5875
DOI
10.1016/j.biocel.2011.07.005
Published
2011-11-00
Epub
2011-00-20
Pages
1563-72
Language
English
Region
Netherlands
NLM ID
9508482
PMCID
PMC3476475
Subset
IM
Grants
NIH HHS · P40 OD011050 · United States
NCRR NIH HHS · P40RR017447 · United States
NIAMS NIH HHS · AR 48323 · United States
NCRR NIH HHS · P40 RR017447 · United States
NIAMS NIH HHS · R01 AR048323 · United States
NCRR NIH HHS · P40 RR017447-03 · United States
NIAMS NIH HHS · AR 47796 · United States
NIAMS NIH HHS · R01 AR044210-04 · United States
External Links
PubMed source DOI: 10.1016/j.biocel.2011.07.005 PMC Full Text View journal details
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