| Project/Area Number |
07671143
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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 |
内分泌・代謝学
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| Research Institution | The University of Tokushima |
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Principal Investigator |
YOSHIMOTO Katsuhiko The University of Tokushima, School of Medicine, Associate Professor, 医学部, 客員助教授 (90201863)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAOKA Takashi The University of Tokushima, School of Medicine, Research Associate, 医学部, 寄付講座教員(助手担 (40263826)
IWAHANA Hiroyuki The University of Tokushima, School of Medicine, Research Associate, 医学部, 寄付講座教員(助手担
板倉 光夫 徳島大学, 医学部, 客員教授 (60134227)
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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 |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1996: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1995: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | thyroid tumors / malignant transformation / mRNA / differential display / AP-PCR / 欠喪 / 増幅 / differential display法 |
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| Research Abstract |
To elucidate the molecular basis for malignant transformation of thyroid tumors, changes in mRNA and genome were analyzed with the differential display method and arbitrarily primed (AP)-PCR method.
1. Differential display method
RNA was extracted from a thyroid follicular adenoma cell line, three thyroid papillary carcinoma cell lines, a thyroid follicular carcinoma cell line, and two undifferentiated thyroid carcinoma cell lines. Bands with different strength of signals among cell lines in the differential display method were marked in the autoradiography and cut out from the gels. They were amplified, cloned, and sequenced. Half of the clones were the known genes such as ribosome RNA,mitochondrial gene, fibronectin, R-ras, a subunit of proteasome, 23 kD highly basic protein. Half of the clones were unknown genes. Northern blot analysis using these clones as probes showed that any clones have the expression levels less than 2-fold among cell lines.
2. AP-PCR method
Genomic DNAs were extracted from 12 thyroid tumor cell lines. DNAs were amplified in conditions that an arbitrarily oligonucleotide with 20 uncleotides was used as a primer and DNA templates were amplified with the low annealing temperature in the first 5 cycles. Patterns of bands among cell lines in the autoradiograph were compared. A loss of signal suggesting deletion was found in a thyroid papillary carcinoma cell line, however, an apparent deletion was not detected in southern blot analysis using the cloned DNA as a probe. In addition, no signals suggesting gene amplification in cell lines were not found.
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