Influence of ADAM28 on biological characteristics of human dental follicle cells.
OBJECTIVE
The aim of this study was to investigate the effects of a disintegrin and metalloproteinase 28 (ADAM28) on the biological characteristics of human dental follicle cells (HDFCs) and possible action mechanism.


METHODS
Eukaryotic expression plasmid containing ADAM28 coding region and ADAM28 antisense oligodeoxynucleotides (AS-ODN) with FITC labelling were constructed and synthesised by gene clone and recombination. Then we respectively transfected them into HDFCs by Lipofectamine 2000 system and detected their effects on proliferation, differentiation and apoptosis of HDFCs by MTT assay, cell cycle detection, ALP activity and Annexin V-FITC/PI analysis. Finally we observed the effects of ADAM28 AS-ODN on HDFCs expressing extracellular matrix (ECM) proteins by immunocytochemical staining.


RESULTS
ADAM28 eukaryotic plasmid was constructed and identified successfully, and could be correctly translated and expressed in HDFCs, furthermore overexpression of ADAM28 promoted the HDFCs proliferation and inhibited specific differentiation of HDFCs, while inhibition of ADAM28 exerted the opposite effects and induced apoptosis. Moreover ADAM28 could significantly inhibit the secretion of OPN and type III collagen of HDFCs.


CONCLUSIONS
ADAM28 might actively participate in the network regulation which associates HDFCs proliferation, differentiation, apoptosis with matrix mineralisation during tooth development by interacting with multiple signal molecules.
Zhao
Zheng
Z
Institute of Stomatology, General Hospital of Chinese People's Liberation Army, 28 Fuxing Road, Beijing 100853, China.
Liu
Hongchen
H
Jin
Yan
Y
Lingling
E
E
D051722:ADAM Proteins / Q000235:genetics / Q000502:physiology*; D000469:Alkaline Phosphatase / Q000032:analysis; D017304:Annexin A5; D017209:Apoptosis / Q000502:physiology; D002453:Cell Cycle / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation; D002648:Child; D024061:Collagen Type III / Q000032:analysis; D004396:Coloring Agents; D003795:Dental Sac / Q000166:cytology* / Q000502:physiology; D019483:Disintegrins / Q000235:genetics / Q000502:physiology*; D004791:Enzyme Inhibitors / Q000032:analysis; D016326:Extracellular Matrix Proteins / Q000032:analysis; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives; D005456:Fluorescent Dyes; D006801:Humans; D058574:Integrin-Binding Sialoprotein; D008562:Membrane Glycoproteins / Q000235:genetics / Q000502:physiology*; D020319:Oligodeoxyribonucleotides, Antisense / Q000235:genetics; D053495:Osteopontin / Q000032:analysis; D010750:Phosphoproteins / Q000032:analysis; D010957:Plasmids / Q000235:genetics; D012795:Sialoglycoproteins / Q000032:analysis; D013778:Tetrazolium Salts; D013844:Thiazoles; D014162:Transfection; D000099311:Dentin Sialophosphoprotein
D051722:ADAM Proteins; D000469:Alkaline Phosphatase; D017304:Annexin A5; D024061:Collagen Type III; D004396:Coloring Agents; D019483:Disintegrins; D004791:Enzyme Inhibitors; D016326:Extracellular Matrix Proteins; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D058574:Integrin-Binding Sialoprotein; D008562:Membrane Glycoproteins; D020319:Oligodeoxyribonucleotides, Antisense; D053495:Osteopontin; D010750:Phosphoproteins; D012795:Sialoglycoproteins; D013778:Tetrazolium Salts; D013844:Thiazoles; D000099311:Dentin Sialophosphoprotein; C492760:FITC-annexin A5; C546871:IBSP protein, human; C410224:ADAM28 protein, human; C022616:thiazolyl blue