| Project/Area Number |
07670240
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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 |
Experimental pathology
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
SHIMADA Toshihide Kyoto Univ.School of Med., Research Associate, 医学研究科, 助手 (30231690)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Rei Kyoto Univ.School of Med., Associate Professor, 医学研究科, 助教授 (60144565)
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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 |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1996: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1995: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | NF1 / small cell lung cancer / differentiaiton / neuroendocrine / splicing / ras |
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| Research Abstract |
Two distinct transcripts, type I and type II,of the neurofibromatosis 1 (MF1) gune are generated by alternative splicing in the region corresponding to the gene's GTP ase-activating protein-related domain (GRD). Relative expression levels of these 2 transcripts were previously correlated to neural differentiation. Since small-cell lung carcinoma (SCLC) often exhibits neuroendocrine properties, we analyzed the type-I to type-II mRNA ratio in 15 SCLC cell lines, using reverse transcriptase and polymerase chain reaction methods. The type-1 mRNA was predominant in 10 cell lines ; 8 of them grew as floating aggregates in culture and had high L-dopa decarboxylase (DDC) activity. The other 5 lines predominantly expressed type-II mRNA,adhered to the culure substrate, and expressed low or undetectable levels of neural cell-adhesion molecule (NCAM) antigen and DDC activity. N2+, one of the subclones of NC1-N417 cells, exhibited a higher type-I to type-II ratio after the cells had adhered to a laminincoated plate and had emitted neurite-like processes.
Using plasmid with Tet-inducible promoter, clones that express after reversion of type I/II ratio were isolated and characterized by the differential RT-PCR method. Since two such clones did not show any homology to know genes, further chatacrerization including sequensing and chromosonal mapping are on going. In case of clinical application, microdissection of frozen sections will be needed for accurate analyasis.
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