| Project/Area Number |
07457331
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | Mie University |
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Principal Investigator |
SUDO Akihiro Mie University Hospital, Lecturer., 医学部・付属病院, 講師 (60196904)
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| Co-Investigator(Kenkyū-buntansha) |
KASAI Yuichi Mie University Hospital, Assistant, 医学部・付属病院, 助手 (20242943)
FUJINAMI Shuichi Mie University Hospital, Assistant, 医学部・付属病院, 助手 (30199351)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥7,400,000 (Direct Cost: ¥7,400,000)
Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1995: ¥6,100,000 (Direct Cost: ¥6,100,000)
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| Keywords | Osteosarcoma / Retinoblastoma / Rb gene / p53 gene / Bone & soft tissue sarcoma / Li-Fraumeni Syndrome / Molecular biology |
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| Research Abstract |
Recently, a continuously growing cell line, designated OSrb/N-M,has been established in vitro from a human osteosarcoma developed in a 17-year-old female patient who had been suffered from bilateral retinoblastoma at infancy. In the previous immunoblot studies with anti-RB and anti-p53 oncoprotein antibodies, the cells were suggested to have mutations at the both gene loci. Thus, we performed an intensive mutation survey of both the Rb and p53 gene in this cell line and the sublines as well as the constitutional cells and the paraffin-embedded retinoblastoma tissue. Any structural anomalies at the Rb gene locus were detected in neither constitutional cells nor osteosarcoma cell lines by Southern blot. However, loss of heterozygosity (L0H) for the RFLPs at introns 1,20 and 25 of Rb gene in osteosarcoma and at the introns 20 and 25 in retinoblastoma were demonstrated byPCR-RFLP analyzes. Furthermore, in contrast to Rb tissue and constitutional cells, homozygous point mutation at the exon 8 of p53 gene was detected in this cell line by PCR-SSCP analysis, and a substitution of guanine for hymine at nucleotide 1480 was identified by sequencing. These results suggest that different second hits might be required for the tumorigenesis of retinoblastoma and osteosarcoma. This is the first report describing the molecular analyzes of Rb and p53 gene mutations of osteosarcoma and retinoblastoma in the same patient with second primary osteosarcoma and bilateral retinoblastoma.
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