チトクロ-ムPー450を中心とした薬物代謝酵素遺伝子発現と化学発癌

1990 年度 実績報告書

• 研究課題/領域番号: 02151006
• 研究機関: 東北大学
• 研究代表者: 渡辺 民朗 東北大学, 抗酸菌病研究所, 教授 (40006101)
• 研究分担者: 山添 康 慶應義塾大学, 医学部, 助教授 (00112699) ; 藤田 正一 北海道大学, 獣医学部, 教授 (10143314) ; 十川 和博 東北大学, 理学部, 助教授 (80175421) ; 佐藤 清美 弘前大学, 医学部, 教授 (50006079) ; 鎌滝 哲也 北海道大学, 薬学部, 教授 (00009177)
• キーワード: チトクロ-ムPー450遺伝子 / チトクロ-ムPー450cDNA / チトクロ-ムPー450蛋白 / 癌原性化学物質の代謝 / 遺伝子転写調節蛋白 / アセチルトランスフェラ-ゼ / グルタチオンSトランスフェラ-ゼ / ヒト遺伝子のRFLPs

研究概要

1.P450、アセチルトランスフェラ-ゼ(AT)の遺伝子構造とその発現:メチルコラントレンで誘導される蟹食いサル肝IA1(ベンツピン代謝)、ハムスタ-肝IA2(芳香族アミン類代謝)、IIA3(アフラトキシン代謝)の、またラット肝培養細胞からP450IICのcDNAを分離した。サル肝IA1、ヒトとビ-グル犬肝のIA2、ヒト肝IIIA4は酵母で、ATはCOS細胞と大腸菌で発現させ、それぞれのcDNAよりの推定アミノ酸配列がそれぞれのたんぱくであることを確かめ、併せて特異的代謝活性の確認も行われた。
2.P450遺伝子の転写調節:ラット肝IA1のXRE(シス転写調節域)に結合するリセプタ-たんぱく(AhR)の部分精製が進行中である。またIA1のプロモ-タ-要素であるBTEに結合するBTEーBのcDNAを分離し、そのアミノ酸組成が転写調節因子Sp1に類似していることを確かめた。ハムスタ-IA1上流域では4ヶのXRE、1ヶのBTEの存在が確かめられた。
3.P450、グルタチオントランスフェラ-ゼ(GT)のたんぱく構造と機能:発癌剤アリルメタノ-ルを不活性化する新しいラット肝GTの分離、ヒト肝からはGTMuの新しい分子種の分離が行われた。ジニトロピレンの不活性化を変異原性テストで調べると、ラットではIA、ヒトではIII4酵素が強かった。
4.ヒトP450遺伝子の多型:白血球DNAを制限酵素で切断するとRFLPが見られ、IA1のMsp1多型、またはIIE1(ジアルキルニトロサミン代謝)のDra1の多型と肺癌発生の感受性との相関が認められた。さらにIIE1の5'上流域にも多型があることが確かめられた。特にIA1のMsp1多型は、喫煙量、扁平上皮癌発生との関連で注目されている。

研究成果

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