| Project/Area Number |
08457435
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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 |
Obstetrics and gynecology
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| Research Institution | Chiba University |
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Principal Investigator |
SEKIYA Souei Chiba University School of Medicine, Obstetrics & Gynecology, Professor, 医学部, 教授 (00092065)
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| Co-Investigator(Kenkyū-buntansha) |
SHIMIZU Kumiko Chiba University School of Medicine, Obstetrics & Gynecology, Assistant, 医学部・付属病院, 助手 (70272317)
OSADA Hisao Chiba University School of Medicine, Obstetrics & Gynecology, Assistant, 医学部, 助手 (30233505)
SUGITA Michio Chiba University School of Medicine, Obstetrics & Gynecology, Lecturer, 医学部・付属病院, 講師 (60196730)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥8,100,000 (Direct Cost: ¥8,100,000)
Fiscal Year 1997: ¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 1996: ¥4,400,000 (Direct Cost: ¥4,400,000)
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| Keywords | Ovarian cancer / Oncogenesis / Telomerase / Telomere / quantitative measurement |
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| Research Abstract |
In this study we focused on the telomerase gene among a group of those associated with oncogenesis of ovarian cancers. By a variety of available molecular and cytogenetic techniques, cell lines and surgical specimens of ovarian cancers have been analyzed to clinically investigate following questions. 1) whether the shortening of telomere length and the acceleration of telomerase activity are correlated to carcinogenesis or not, 2) if so, to which stage in tumor progression, 3) how they communicate with other oncogenes and tumor suppresser genes.
We designed new PCR primers for detecting telomerase activity, which are different from ordinary ones used in TRAP assay and succeeded in performing electrophoresis and computer analysis on an autosequencer using theses fluorescence-labeled primers. Fragment analysis program led to accurate and convenient quantitative measurement of fluorescent ladder bands produced by telomerase. This newly developed assay system was applied to quantitative ana
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lysis of telomerase activity in cell lines, primary or metastatic tumors, recurrence tumors after chemotherapy, and formalin-fixed tissue sections.
In 6 ovarian cancer cell lines we examined high degrees of telomerase activity were detected as expected. However, we failed to obtain stable results for surgical and pathological specimen of ovarian cancers. This failure was thought to result from some inhibitory substance for telomerase or PCR reaction in tissue extracts. Several extraction techniques for tissue proteins were thus examined, but this problem has not been overcome in the research period. We also developed in situ detection of telomerase activity in smear cells of ovarian and cervical cancers, and obtain preliminary results promising clinical application.
It is scheduled to make continuous improvements to establish quantitative measurement of telomerase activity in povarian cancers and, compare degrees of their activity with clinical background including stage, tissue type and grade, and expression of a panel of oncogenes and cell cycle-related genes. Our final goal is to clarify the roles of telomerase in oncogenesis mechanism and examine the ossibility of its clinical application. Less
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