2001 Fiscal Year Final Research Report Summary
Investigation of oral cancer development by using apoptosis and its application for clinical evaluation
| Project/Area Number |
12671836
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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 |
病態科学系歯学(含放射線系歯学)
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| Research Institution | Kyushu Dental College |
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Principal Investigator |
OHBA Takeshi Kyushu Dental College, Professor, 歯学部, 教授 (10047798)
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| Co-Investigator(Kenkyū-buntansha) |
UEYAMA Yoshiya Tottori University, School of Medicine, Associate Professor, 歯学部, 助教授 (00168668)
HANEJI Tatsuji Tokushima University, School of Dentistry, Professor, 歯学部, 教授 (50156379)
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| Project Period (FY) |
2000 – 2001
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| Keywords | Anti-canccer drug / Apoptosis / Egr-1 / NORs / Oral carcinoma / Protein phosphatase |
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| Research Abstract |
Oral squamous carcinoma cell line SSCKN cells were shown to be highly sensitive to bleomycin, whereas SCCKN cells were minimally sensitive to this reagent. Peplomycin caused apoptosis SCCKN and SCCTF cells in a dose-dependent fashion with the maximal effect at concentrations of 5 μM and 50 μM, respecnvely, as determined by phase-contrast microscopy, WST-1 cell viability assay, Hoechst 33342 staining, and DNA ladder formation. These results indicate that peplomycin-induced apoptosis in oral squamous carcinoma cell lines depends on the sensitivity of these cells to bleomycin.
We examined the expression of Egr-1 in human squamous carcinoma cell line SCCKN cells treated with okada1C acid. Western blot analysis revealed that Egr-1 was strongly expressed in SCCKN cells and that okadaic acid decreased the expression of Egr-1 protein in these cells. Suppression of Egr-1 protein expression okadaic acid-treated cells stemmed from the suppression of the Egr-1 mRNA level, as determined by the RT-RCR method. A gel mobility shift assay showed that treatment of SCCKN cells with okadaic acid suppressed Egr-1 binding to the DIG-labeled Egr-1 consensus oligonucleotide probe. Treatment of SCCKN cells with okadaic acid suppressed the cell viability of SCCKN cells and induced apoptosis in these cells.
The expression and cytolocalization of protein phosphatase type 1 δ isoform and nucleolin in human osteoblastic MG63 and Saos-2 cells. Anti-nucleolin antibody interacted with the 100- and 95-kDa proteins present in the whole cell lysate. The 100-kDa protein was detected in nuclear and nucleolar fractions. The 95-kDa protein was detected in cytosolic and nucleoplasmic fractions. Protein phosphatase 1δ and nucleolin were co-localized and interact with each other in nucleolus in MG63 and Saos-2 cells revealed bv immunofluorescent and immunoprecipitation method.
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