2004 Fiscal Year Final Research Report Summary
Control of osteoclast differentiation by using siRNA for ICAM-1 gene silencing
| Project/Area Number |
15591939
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Morphological basic dentistry
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| Research Institution | KYUSHU UNIVERSITY |
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Principal Investigator |
NAGATA Kengo Kyushu University, Faculty of Dental Science, Research Associate, 大学院・歯学研究院, 助手 (90189134)
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| Co-Investigator(Kenkyū-buntansha) |
IIJIMA Tadahiko Kyushu University, Faculty of Dental Science, Professor, 大学院・歯学研究院, 教授 (50090874)
KUKITA Toshio Kyushu University, Faculty of Dental Science, Associate Professor, 大学院・歯学研究院, 助教授 (70150464)
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| Project Period (FY) |
2003 – 2004
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| Keywords | RNAi / osteoclast / ICAM-1 / LFA-1 / cell fusion |
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| Research Abstract |
To study the involvement of cell adhesion molecules in the osteoclastogenesis, we examined the expression of intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1) on the macrophage cell line RAW D clone which effectively differentiated into multinucleated osteoclast-like cells in the α-MEM with RANKL and TNF-α. The ratios of the number of ICAM-1-positive mononuclear cells to total mononuclear cells were enhanced by the addition of RANKL and TNF-α in the α-MEM, however, the ratios of the number of LFA-1-positive mononuclear cells to total mononuclear cells were the same in the α-MEM with/without RANKL and TNF-α on day 1 in culture. Although ICAM-1 was expressed on mononuclear cells and osteoclast-like multinucleated cells in the α-MEM with RANKL and TNF-α, LFA-1 expression was limited on mononuclear cells in the α-MEM with/without RANKL and TNF-α on day 2 in culture. By immunofluorescent method, the same mononuclear cell which was deemed preosteoclast was positive for both ICAM-1 and LFA-1 on day 2 in culture. To confirm the involvement of ICAM-1 in the osteoclastogenesis, we knocked down the ICAM-1 mRNA expression by using siRNA for ICAM-1 in the RAW D cell culture. ICAM-1 gene silencing inhibits the multinucleated osteoclast-like cell formation derived from RAW D clone in the α-MEM with RANKL and TNF-α after day 3 in culture. These results suggest that the firm adhesion by the interaction of ICAM-1/LFA-1 among preosteoclasts stimulates the formation of multinucleated osteoclasts.
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