| Project/Area Number |
05454395
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Cerebral neurosurgery
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| Research Institution | Kanazawa University |
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Principal Investigator |
YAMASHITA Junkoh Kanazawa University professor, 医学部, 教授 (90026948)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAGUCHI Kazuo 99999999, 遺伝子実験施設, 施設長教授 (00019879)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1994: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1993: ¥3,600,000 (Direct Cost: ¥3,600,000)
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| Keywords | DNA triplex, antigene, EGFR,Sp-1, human glioma, human squamous cell carcinoma, retrovirus vector, gene therapy / アンチジーン / 上皮成長因子受容体 / 癌遺伝子 / グリオーマ / 扁平上皮癌 / レトロウイルスベクター / 遺伝子療法 / 三重鎖DNA / triple stranded DNA / anti-gene strategy / EGFR / Sp-1 / glioma |
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| Research Abstract |
Mixed purine-pyrimidine anti-gene oligodeoxynucleotides were designed to form collinear DNA triplexes with pyrimidine-rich elements in the human epidermal growth factor receptor (EGFR) gene promoter as gene code blocker. Their effects as suppressors of the EGFR gene transcription were evaluated using human squamous cell carcinoma (A431) and human glioma (U251MG and U87MG) cell lines. Gel shift analyzes indicated that the oligonucleotide forms a collinear triplex within the Sp-1 binding site. An in vitro assay revealed a correlation between the triplex formation and the suppression of EGFR transcription. We postulate that guanine residues are not always optimum in apposition to G-C pairs to form triple helices in the target. We found that oligonucleotides designed to form a triple helix with enhancer elements of the EGFR gene promoter can suppress mRNA formation and also the proliferation of a human glioma cell line. Anti-gene binding to a DNA duplex may serve as a basis for an alternative program of gene control in vitro. We are trying to develope retrovirus vectors which allow expression of anti-genes. We also consider the feasibility of an anti-gene strategy as adjuvant therapy of glioma.
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