Semi-quantitative RT-PCR analysis of LIM mineralization protein 1 and its associated molecules in cultured human dental pulp cells.
OBJECTIVE
LIM mineralization protein 1 (LMP-1), an intracellular signaling molecule, regulates osteoblast differentiation and maturation, as well as bone formation. However, the role of LMP-1 in the differentiation of human dental pulp cells and formation of dentin has not been determined. The study was to investigate the expression of LMP-1, the related proteins, such as bone morphogenetic proteins 2, 6 and 7 (BMP-2, BMP-6 and BMP-7), and core binding factor alpha 1 (Cbfa1) during the differentiation of cultured human dental pulp cells and the formation of mineralized nodules.


METHODS
Differentiation of human dental pulp cells was induced by dexamethasone, asorbic acid and beta-glycerophosphate. The formation of mineralized nodules, was determined by Von Kossa staining and immunocytochemistry detection of dentin sialoprotein. Expression of LMP-1, the related proteins and the differentiation marker alkaline phosphatase (ALP) was analysed by reverse transcriptase-polymerase chain reaction (RT-PCR).


RESULTS
The expression of LMP-1, BMP-2, BMP-6, BMP-7 and Cbfa1 was significantly increased in the process of dental pulp cells differentiation and the formation of mineralized nodules, while the pattern of the expression was distinct.


CONCLUSIONS
The elevated level of LMP-1, BMPs and Cbfa1 expression indicated they might play a role in the differentiation of human dental pulp cells and the formation of mineralized nodules.
Zhang
Qi
Q
Key Laboratory for Oral Biomedical Engineering of Ministry of Education and Department of Cariology & Endodontics, Luoyu Road 237, School and Hospital of Stomatology, Wuhan University, Wuhan, Hubei Province 430079, PR China.
Wang
Xiaoying
X
Chen
Zhuo
Z
Liu
Gang
G
Chen
Zhi
Z
D048868:Adaptor Proteins, Signal Transducing; D000293:Adolescent; D000328:Adult; D000469:Alkaline Phosphatase / Q000032:analysis; D000975:Antioxidants / Q000494:pharmacology; D001205:Ascorbic Acid / Q000494:pharmacology; D015415:Biomarkers / Q000032:analysis; D055396:Bone Morphogenetic Protein 2; D055418:Bone Morphogenetic Protein 6; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins / Q000032:analysis; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D050659:Core Binding Factor Alpha 1 Subunit / Q000032:analysis; D003598:Cytoskeletal Proteins; D003782:Dental Pulp / Q000166:cytology*; D003907:Dexamethasone / Q000494:pharmacology; D016326:Extracellular Matrix Proteins / Q000032:analysis; D005938:Glucocorticoids / Q000494:pharmacology; D005994:Glycerophosphates / Q000494:pharmacology; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000032:analysis*; D060588:LIM Domain Proteins; D010750:Phosphoproteins / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012795:Sialoglycoproteins / Q000032:analysis; D014074:Tooth Calcification / Q000502:physiology; D016212:Transforming Growth Factor beta / Q000032:analysis; D016335:Zinc Fingers; D000099311:Dentin Sialophosphoprotein
D048868:Adaptor Proteins, Signal Transducing; D000469:Alkaline Phosphatase; D000975:Antioxidants; D001205:Ascorbic Acid; D015415:Biomarkers; D055396:Bone Morphogenetic Protein 2; D055418:Bone Morphogenetic Protein 6; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins; D050659:Core Binding Factor Alpha 1 Subunit; D003598:Cytoskeletal Proteins; D003907:Dexamethasone; D016326:Extracellular Matrix Proteins; D005938:Glucocorticoids; D005994:Glycerophosphates; D047908:Intracellular Signaling Peptides and Proteins; D060588:LIM Domain Proteins; D010750:Phosphoproteins; D012795:Sialoglycoproteins; D016212:Transforming Growth Factor beta; D000099311:Dentin Sialophosphoprotein; C525718:BMP2 protein, human; C525816:BMP6 protein, human; C525819:BMP7 protein, human; C484380:PDLIM7 protein, human; C031463:beta-glycerophosphoric acid