p53癌抑制遺伝子によるヒト発がん抑制の分子機構

• Project/Area Number: 08264104
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: 土田 信夫 東京医科歯科大学, 歯学部, 教授 (60089951)
• Co-Investigator(Kenkyū-buntansha): 山下 利春 札幌医科大学, 癌研究所, 講師 (50167706) ; 石田 誠一 癌研究会癌研究所, 遺伝子, 研究員 (10270505) ; 高橋 玲 京都大学, 大学院・医学研究科, 助教授 (60144565) ; 山泉 克 熊本大学, 医学部, 教授 (70107093) ; 横田 淳 国立がんセンター研, 生物学部, 部長 (10191503)
• Project Period (FY): 1996
• Project Status: Completed (Fiscal Year 1996)
• Budget Amount: ¥12,500,000 (Direct Cost: ¥12,500,000); Fiscal Year 1996: ¥12,500,000 (Direct Cost: ¥12,500,000)
• Keywords: p53 / 癌抑制遺伝子 / G1停止 / アポトーシス / 熱ショック / HBV / テトラサイクリンプロモーター / 温度感受性変異株

Research Abstract

ヒト癌の半数以上で遺伝子変異検出されているp53癌抑制遺伝子について、細胞DNAへの傷害あるいはストレスがp53の活性化/安定化をもたらす過程(上流シグナル)、および活性化/安定化されたp53からG1停止、アポトーシスへの過程(下流シグナル)を解析し以下の結果を得た。
1.熱ショックストレス後のp53の核での蓄積は翻訳レベルで生じていた(山泉)。2.コドン273をロイシンに変えた人工変異株は転写活性化能を欠くが細胞増殖抑制能を保持していた(山下)。3.テトラサイクリンで野性型p53、またはRbの発現誘導できる細胞株を樹立した。低レベルのp53発現ではG1停止を、高レベルではアポトーシスを誘導できた。一方Rbを骨肉腫細胞株に発現させた場合、分化マーカーの発現がみられた(横田)。4.Fasによってアポトーシス誘導可能な野性型p53発現大腸癌細胞株、および細胞死をともなった分化誘導可能な野性型p53発現口腔扁平上皮癌細胞株を樹立した(高橋)。5.温度感受性p53変異株導入細胞をもちいて解析したところ、Rb発現を欠くがん細胞のG1停止ではp130Rbファミリー蛋白質が重要であった。アポトーシスが誘導される細胞系ではp21に比べ高いMDM2の発現がみられこの比が重要であった(土田)。6.HBVのX蛋白質はp53と結合することなしにp53を細胞質へ移行させp53機能を抑制させる可能性が示された(石田)。なお1996年7月に班会議を1997年3月には外国からの演者を3人加え英語にて公開のシンポジウムを開いた。また班員の所持するp53関連資材の情報交換を行い班員相互の研究がスムースに行える様にした。

Research Products

• [Publications] Munirajan A K,Tsutsumi-Ishii Y,Mohanprasad B K,Hirano Y,Munakata N,Shanmugam G & Tsuchida N.: "p53 gene mutations in oral carcinomas from India." Int.J.Cancer. 66. 297-300 (1996)
• [Publications] Takada S,Kaneniwa N,Tsuchida N & Koike K.: "Hepatitis B virus X gene expression is activated by X protein but repressed by p53 tumor suppresor gene product in the transient expression system." Virology. 216. 80-89 (1996)
• [Publications] Komiyama T,Matsumura K & Tsuchida N.: "Comparison between DNA copy number in original oral squamous cell carcinomas and their corresponding cel lines by comparative genomic hybridization." Jpn.J.Cancer Res.in press.
• [Publications] Adachi J,Ookawa K,Shiseki M,Okazaki T,Tsuchida S,Morishita K,& Yokota J.: "Induction of apoptosis but not G1 arrest by expression of the wild-type p53 gene in small-cell lung carcinoma." Cell Growth & Differentiation. 7. 879-886 (1996)
• [Publications] Shiseki M,Kohno T,Adachi J,Okazaki T,Ohtsuka T,Mizoguchi H,Noguchi M,Hirohashi S,& Yokota J.: "Comparative allelotype of early and advacned stage non-small cell lung carcinomas." Genes,Chromosomes & Cancer. 17. 71-77 (1996)
• [Publications] Yokota J.Ookawa K,Shiseki M,Adachi J,& Ishimaru G.: "Application of molecular genetic information for the diagnosis of colorectal carcinoma.in ″New Strategy for Treatment of Hereditary Colorectal Cancer." (ed.by S.Baba)Churchill Livingstone,Tokyo. 70-79 (1996)
• [Publications] Hui A-M,Sakamono M,Kanai Y,Ino Y,Gotoh M,Yokota J.& Hirohashi S.: "Inactivation of p16ink4 in hepatocellular carcinoma." Hepatology. 24. 575-579 (1996)
• [Publications] Yokota J.Adachi J,Sasaki A & Kohno T.: "Genetic defects in cancer cells and cancer prone families." European J.Cancer Prevention. 5. 33-36 (1996)
• [Publications] Ohtani-Fujita N,Dryja T F,Rapaport J Matsumura T,Ohnishi Y,Hotta Y,Takahashi R,Kato M V,Ishizaki K,Sasaki M S,Hortemke: "Hypermethylation in the retinoblastoma gene is associated with unilateral,sporadic retinoblastoma." Cancer Genet.& Cytogenet.,in press.
• [Publications] Fujita M,Takahashi R.Kitada K,Watanabe R,Kitazawa S,Ashoori F,Liang P,Saya H,Serikawa T,& Maeda S.: "Alterantive splicing of the erythropoietin receptor gene correlates with erythroid differentiation in rat hematopoietic and leukemic cells." Cancer Letter in press.
• [Publications] Ishida S.Takada S & Koike K.: "Isolation and anlysis of cellular DNA fragments derectly binding to c-Myc protein." Leukemia Res.in press.
• [Publications] Koike K,Takada S,Ishida S,Nakanishi M.,Hayashi Y & Kobayashi M.: "in ″Hepatitis B virus : Molecular mechanisms in disease and novel strategies for therapy″." Imperial College Press,London. in press.
• [Publications] Kawamura M,Yamashita T,Segawa K,KaneuchiM,Shindoh M,& Fujinaga K.: "The 273rd codon mutants of p53 show growth modulation activities not correlated with p53-specific transactivation activity." Oncogene. 12. 2361-2367 (1996)
• [Publications] Funaoka K,Shindoh M,Yamashita T,Funinaga K,Amemiya A & Fujinaga K.: "High-resk HPV-positive human cancer cell lines show different sinsitivity to cisplatin-induced apoptosis correlated with the p21wasf1/cip1 level.21GC14:Cancer Lett." 108. 15-23 (1996)
• [Publications] Adachi J,Kohno T.Baeg G-H,Akiyama T & Yokota J.: "Growth suppression of non-small cell lung carcinoma cells by the introduction of the p16ink4 gene." Int.J.Oncol.10. 33-39 (1996)
• [Publications] Yokota J & Harris C C.: "Mutational analysis of coding sequences of the entire transforming growth factor bata type II receptor gene in sporadic human colorectal cancer using genomic DNA and intron primers." Oncogene in press.
• [Publications] Ookawa K,Tsuchida S,Adachi J & Yokota J.: "Differentiation induced by RB expression and apoptosis induced by p53 expression in an osteosarcoma cell line." Oncogene in press.
• [Publications] Tani M,Takenoshita S,Kohno T,Hagiwara K,Nagamachi Y,Harris C C & Yokota J.: "Infrequent mutations of the transforming growth factor beta type II receptor gene at chromosome 3p22 in human lung cancers with chromosome 3p deletions." Carcinogenesis in press.
• [Publications] Takita J.Hayashi Y,Kohno T,Yamaguchi N,Hanada R,Yamamoto K and Yokota J.: "Deletion map of chromosome 9 and p16(CDKN2A) gene alterations in neuroblastoma." Cancer Res.in press.
• [Publications] Ashoori F,Suzuki S,Zhou J,Nishigaki I,& Takahashi R.: "Possible contributions of mastocytosis,apoptosis and hydrolysis in pathophysiology of radomized skin flaps in human and guinea pig." Plastic Reconstr.Surg.98. 491-501 (1996)
• [Publications] Valente P,Melchiori A,Paggi M G,Masiello L,Rabatti D,Santi L,Takahashi R, Albini A,& Noonan D M.: "RB1 oncosuppressor gene over-expression inhibits tumor progression and induces melanogenesis in metastatic melanoma cells." Oncogene. 13. 1169-1178 (1996)
• [Publications] Takahashi R,Inagaki T,MatsuwariS,Fujioka M,Maeda S,Ijuhin N,Haga H,Koh T,Shimada K & Saya H.: "Transfection of wild-type p53 indces differentiation in human gingival carcinoma cells." Eur.J Cancer. 32A. 533-539 (1996)