Molecular biological study on the osteoblast metabolism by EGF.
Grant-in-Aid for General Scientific Research (B)
|Allocation Type||Single-year Grants |
Functional basic dentistry
|Research Institution||Nihon University |
TAKIGUCHI Hisashi Nihon University, School of Dentistry at Matsudo, Professor, 松戸歯学部, 教授 (00050013)
SHIBATA Yasuko Nihon University, School of Dentistry at Matsudo, Instructor, 松戸歯学部, 助手 (90133438)
MORIYA Yoshiko Nihon University, School of Dentistry at Matsudo, Associate Professor, 松戸歯学部, 助教授 (40050017)
ABIKO Yoshimitsu Nihon University, School of Dentistry at Matsudo, Associate Professor, 松戸歯学部, 助教授 (70050086)
TAKIGUCHI Hisashi Nihon University, School of Dentistry at Matsudo, Professor (00050013)
|Project Period (FY)
1988 – 1989
Completed (Fiscal Year 1989)
|Budget Amount *help
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1989: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1988: ¥5,600,000 (Direct Cost: ¥5,600,000)
|Keywords||Osteoblast / Prostaglandin E_2 / EGF / Gene expression / Bone metabolisms / EGF受容体 / 1α / 25VD_3受容体 / collagen / alkaline phosphatase / collagenase inhibitor / m-RNA / DNA probe|
The mechanisms for regulation of prostaglandin E_2 production from the osteoblast cell line MC3T3-El during mineralization and differentiation processes by EGF or cytokines were studied.Cell were challenged with EGF or cytokine at different culture period and harvested, then RNA fractions were isolated. The gene expression was analyzed as MRNA transcription rate using Northern-hybridization. Results were as follows;
1. In culture of MC3T3-El cells over a 28-d period, alkaline phosphatase activity was increased by 10- and 20-day and mineralization was started by 20-day. These data suggest that 10- and 20-day may be an important stage for differentiation as bone cell
2. Northern blot hybridization data using several synthetic DNA probe show
(1). The mRNA levels for type I collagen decreased after 21-day period.
(2). The expression rate of cyclooxygenase gene was similar manner to that of prostaglandin E2 production from 7- to 28-day period.
(3). The mRNA levels for osteonectin were from 7-day and then decreased to 21-day period when mineralization started.
(4). The expression of collagenase inhibitor (TIMP) gene was stimulated by TGF-beta and suppressed by platelet activating factor (PAF).
(5). The expression of TIMP gene in RCJ-3.1 cell line was increased during the passage of culture, mRNA level of TIMP was kept constant during culture periods in RCB-2.2-cells.
3. In MC3T3-El cells, PAF stimulated the prostaglandin E_2 production in time and dose dependent manner but PAF did not stimulated the arachldonic acid release. The stimulation of prostaglandin E_2 production was reduced by the pretreatment of the cells with a PAF antagonist. Furthermore.
4. The release of interleukin-6 form human ligament cells was significantly stimulated by interleukin-1alpha , tumor necrosis factor-alpha and E.coli LPS.
Report (3 results)
Research Products (15 results)