| Project/Area Number |
06671854
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Functional basic dentistry
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| Research Institution | Okayama University |
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Principal Investigator |
TAKAHASHI Kojiro Okayama University, Dental School, Associate Professor, 歯学部, 助教授 (00144775)
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| Co-Investigator(Kenkyū-buntansha) |
TAKIGAWA Masaharu Okayama University, Dental School, Professor, 歯学部, 教授 (20112063)
HATTORI Takako Okayama University, Dental School, Assistant Professor, 歯学部, 助手 (00228488)
NAKANISHI Tohru Okayama University, Dental School, Assistant Professor, 歯学部, 助手 (30243463)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1995: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1994: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | Insulin-like growth factor II / Insulin-like growth factor I / Osteoblastic cell / Chondrosarcoma / Chondrocytes / Transcriptional promoter / RT-PCR / RNase Protection Assay / Insulin-like growth factor-II / Insulim-like growth factor-I / 軟骨様細胞 / 転写調節 / Insulin-like growth factor-I |
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| Research Abstract |
Using a chondrocytic cell line HCS-2/8 derived from human chondrosarcoma, the quantitative analytical method adapting the RNase protection Assay (RPA) was developped in order to examine the expression and promoter-alternation of insulin-like growth factor genes. The probe of the RPA experiments was constructed for the promoters 2P-1,2P-3 and 2P-4 of IGF-2 and the promoters 1P-1 and 1P-2 of IGF-1, but the promoter 2P-2 of IGF-2 failed to be constructed because the transcriptional product was very little in HCS-2/8. Now, we continue to investigate the determination of detectable limitation of the RPA method using their probes, and we will retry to construct the probe asfter the accumulation of RT-PCR product for 2P-2 transcript.
During the present developmental investigations, we found some new facts as following :
1.The simultaneous expression of four promoters of IGF-2 were induced in HCS-2/8 and human normal chondrocytes.
2.A defect of four base pair in the promoter 2P-4 transcript of IGF-2 in HCS-2/8 was observed at the 3'-end of exon 6, at which the position is upstream of 9 to 6 base from the translation-starting point. This defect may be due to the alternative splicing specific to the chondrosarcoma.
3.Comparing with HCS-2/8 and human normal chondrocytes, we found a parallel correlation between IGF-2 and H19 (tumor reppressor gene) expressions.
4.The restriction fragment length polymorphism of IGF-2 gene in HCS-2/8 suggested that only the paternal allele (s) existed at the genomic level, but the maternal imprinting allele was absent in the chondrosarcoma. This perhaps correlate with the tumor formation.
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