MiR-143 and miR-135 inhibitors treatment induces skeletal myogenic differentiation of human adult dental pulp stem cells.
OBJECTIVE
Dental pulp stem cells (DPSCs) possess pluripotent properties that allow them to differentiate into multiple cell lineages, which can be potentially used in tissue regeneration. The aim of this in vitro study is to explore the effect of miRNAs on the myogenic differentiation of human adult DPSCs and seek for some potential biological factors for stable and feasible application in DPSC myogenic differentiation.


METHODS
Human adult DPSCs were isolated from normal impacted third molars were treated with 5-Aza-2'-deoxycytidine to induce to myogenic differentiation in vitro. During this process the levels of myomiRNAs and myogenic marker genes were detected by real-time qPCR and Western blotting. Then antisense oligonucleotides of miR-143 and miR-135 were transfected into DPSCs to explore their effects on myogenic differentiation. Gene expression detection and MyHC immunofluorescence microscopy analysis were applied to characterize the myogenic differentiation of DPSCs.


RESULTS
Expression of miR-135 and miR-143 was markedly decreased in myoblast DPSCs induced by 5-Aza. Part of the DPSCs treated with miR-135 or miR-143 inhibitors showed apparent myocytic properties and eventually fused to form myotubes. Co-transfection of miR-135 and miR-143 inhibitors impelled half of DPSCs to form myotubes.


CONCLUSIONS
MiR-135 and miR-143 inhibitors could induce myogenic differentiation of DPSCs. Our findings indicated that miRNAs could exert a decisive function in induction of myogenic differentiation of DPSCs.
Department of Stomatology, The General Hospital of the Air Force, Beijing, China. Electronic address: dongxiali2014@163.com.
Dongxia
D
Li
Department of Stomatology, The General Hospital of the Air Force, Beijing, China.
Tianzheng
T
Deng
Department of Stomatology, The General Hospital of the Air Force, Beijing, China.
Hongshi
H
Li
Department of Stomatology, The General Hospital of the Air Force, Beijing, China.
Ying
Y
Li
D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D001374:Azacitidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002478:Cells, Cultured; D000077209:Decitabine; D003782:Dental Pulp / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D035683:MicroRNAs / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics; D008964:Molar, Third / Q000166:cytology / Q000187:drug effects; D024510:Muscle Development / Q000187:drug effects* / Q000235:genetics; D018482:Muscle, Skeletal / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D032446:Myoblasts / Q000166:cytology / Q000502:physiology; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000235:genetics; D014162:Transfection; D055815:Young Adult
C544965:MIRN135 microRNA, human; C542359:MIRN143 microRNA, human; D035683:MicroRNAs; D016376:Oligonucleotides, Antisense; D000077209:Decitabine; D001374:Azacitidine
Antisense oligonucleotide inhibitor; Dental pulp stem cell; Myogenic differentiation; miR-135; miR-143