2001 Fiscal Year Final Research Report Summary
Interaction between signaling molecules and HSP90 on apoptosis pathway in HSG cells
| Project/Area Number |
12671813
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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 |
Functional basic dentistry
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| Research Institution | Iwate Medical University |
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Principal Investigator |
SATO Nobuko Iwate med. Uni. Sch. Dent., Dept. Biochem., Professor, 歯学部, 教授 (00048399)
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| Co-Investigator(Kenkyū-buntansha) |
CHOSA Naoyuki Iwate med. Uni. Sch. Dent., Dept. Biochem., Research Associate, 歯学部, 助手 (80326694)
KYAKUMOTO Seiko Iwate med. Uni. Sch. Dent., Dept. Biochem., Assitant Professor, 歯学部, 講師 (90118274)
KAMO Masaharu Iwate med. Uni. Sch. Dent., Dept. Biochem., Associate Professor, 歯学部, 助教授 (40214564)
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| Project Period (FY) |
2000 – 2001
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| Keywords | Apoptosis / HSP90 / Fas / geldanamycin / salivary gland adenocarcinoma / caspase / TNF-α |
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| Research Abstract |
Recently, it has been reported that a number of signaling molecules are regulated by the HSP family proteins which function as molecular chaperones. The present study focused on the interaction between signaling molecules and HSP90 on apoptosis pathway in the human salivary gland adenocarcinoma cell line (HSG).
In HSG cells, TNF-α alone did not induce apoptosis. On the other hand, agonistic Fas antibody alone induced apoptosis in 30 % of the cells and TNF-α treatment followed by agonistic Fas antibody induced apoptosis in 60 % of the cells. RT-PCR analysis of apoptotic signaling molecules after TNF-α treatment revealed the highly expression of Fas. Thus, it was suggested that Fas increased by TNF-α enhanced the agonistic Fas antibody-mediated apoptosis. From the results of the experiment with inhibitors for caspases which play important roles for apoptotic pathway, both caspase-8 and caspase-3 are involved in the agonistic Fas antibody-induced apoptosis.
HSG cells were treated with geldanamycin which is a specific inhibitor for HSP90. While agonistic Fas antibody showed apoptosis in 30 % of the cells, the combination of geldanamycin and agonistic Fas antibody increased to 90 % of the cells. This result suggested that HSP90 participates in the regulation of apoptotic signaling molecules. Western blotting analysis after the agonistic Fas antibody stimulation showed that the protein level of HSP90β isoform increased while that of HSP90α did not changed. Furthermore, proteome analysis of HSG cells was performed. Protein spots of HSP90 observed on the two-dimensional gel electrophoresis were shifted to the more acidic after the agonistic Fas antibody stimulation, suggesting the phosphorylation of HSP90.
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Research Products
(10 results)
