1998 Fiscal Year Final Research Report Summary
Establishment of new method for cancer diagnosis : Telomerase and human telomerase RNA component
| Project/Area Number |
09671239
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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 |
General surgery
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| Research Institution | Sapporo Medical University School of Medicine |
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Principal Investigator |
OHMURA Tousei Department of Surgery Sapporo Medical University, School of Medicine, Instructor, 医学部, 助手 (30295349)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Naoki Sapporo Medical University, School of Medicine, Professor, 医学部, 教授 (10158644)
HIRATA Koichi Sapporo Medical University, School of Medicine, Professor, 医学部, 教授 (50136959)
HATA Fumitake Sapporo Medical University, School of Medicine, Instructor, 医学部, 助手 (70291557)
YAGIHASHI Atsuhito Sapporo Medical University, School of Medicine, Assistant Professor, 医学部, 講師 (40260757)
OKAZAKI Akira Sapporo Medical University, School of Medicine, Assistant Professor, 医学部, 講師 (50213921)
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| Project Period (FY) |
1997 – 1998
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| Keywords | テロメラーゼ / TRAP法 / テロメラーゼ関連遺伝子 |
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| Research Abstract |
We established non-radioisotopic TRAP assay using Digoxigenin labeled probe and TaqMan RT-PCR for hTR.Using this assay, we quantified hTR expression in two human pancreatic cancer cell lines, ASPC-1 and MIAPaCa-2. Our results indicated that hTR expression in MIAIPaCa-2 was 1.99-fold higher than that in ASPC-1 cells. This TaqMan RT-PCR assay appears to be useful in determining the amount of hTR in clinical specimens.
We have attempted to detect telomerase activity by Southern bolt analysis using a digoxigenin labeled probe (5'-(CCCTTA)6CCCTAA-3'). Telomerase activity was detected in all cancer cell lines that were tested, and : the lower limit for detection was 10^3 cells. Although telomerase activity was detected in colon cancer tissues. It was not found in adenoma or normal tissues. Telomerase activity was completely abrogated by heat and RNase treatment of the CHAPS extract. Our results demonstrate that this nonradioisotopic assay is safe and reliable for detecting telomerase activity in cancer cell lines and biopsy samples.
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