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頭頸部扁平上皮癌における遺伝子変異と蛋白表現型によるp53機能分類

小林, 謙也 東京大学 DOI:10.15083/0002004178

2022.06.22

概要

【背景】
 TP53変異は頭頸部扁平上皮癌において最も高頻度に出現する変異であり、予後と密接に関与しているが、日常診療において予後因子として使用されていない。この理由として、TP53変異がp53蛋白に与える影響が多様であることが挙げられる。ナンセンス変異、フレームシフト変異、スプライシング異常は単独でp53蛋白の機能不全を引き起こすが、ミスセンス変異は変異部位、アミノ酸置換の種類により、p53蛋白に及ぼす影響は様々である。さらに、DNAはそれ自体に機能があるわけではなく、RNAに転写、蛋白に翻訳され初めて機能が生じる。現在、次世代シーケンサーを用いたクリニカルシーケンスはがん遺伝子解析の中心となりつつあるが、ゲノム情報に加え、蛋白レベルも考慮した遺伝子機能評価が重要であると考えられる。
 頭頸部扁平上皮癌の外科治療では肉眼病変を超えた癌の浸潤や異形上皮を同定するために、ルゴール染色や術中迅速病理検査も活用しつつ、病理学的完全切除を狙う。しかしながら実臨床では、断端陰性であっても、TP53変異を伴う前癌病変から局所再発を来すことがある。従来の術前・術中検査による切除範囲の決定には限界があると考えられ、ゲノム情報に基づく腫瘍の分子生物学的な特徴に合わせた切除断端距離の設定も重要課題である。
 本研究は、頭頸部扁平上皮癌の初回腫瘍を次世代シーケンサー及び免疫染色を用いてTP53遺伝子を中心に解析することにより、TP53変異とp53蛋白表現型の併用解析が精度の高い臨床予後予測を可能にし、個別化治療に反映できるか検討することを目的とした。

【方法】
 2013年1月から2015年12月までに国立がん研究センター中央病院頭頸部外科で外科的治療された317例の初回腫瘍のホルマリン固定パラフィン包埋(FFPE)検体を用いて、次世代シーケンサーによるTP53、PIK3CA、AKT、HRAS遺伝子変異解析、及び免疫染色によるp53、p16蛋白表現型解析を施行した。さらに、TP53変異型とp53蛋白表現型を併用解析することでp53機能分類を行った。変異型TP53でかつp53蛋白表現型がExtremely positive(EP)、Lostであるものをp53機能不良群、野生型TP53または変異型TP53でかつp53蛋白表現型がOthersであるものをp53機能良好群と分類した。このp53機能分類と粗生存、局所無再発生存、遠隔無再発生存との相関や、外科的切除断端距離別の局所予後との相関を解析した。

【結果】
 p16陽性中咽頭癌はその他の頭頸部癌と比較して統計学的に有意に若年者に多く、Smokerの頻度が少なく、粗生存率が高かった。しかしながら、TP53変異の有無で粗生存率に統計学な差を認めなかった。
 p16陽性中咽頭癌を除外した頭頸部扁平上皮癌284例の内、p53機能不良群は140例、p53機能良好群は144例であり、p53機能分類は粗生存、局所無再発生存、遠隔無再発生存に対して独立した予後因子であった。PIK3CA、AKT、HRAS変異は予後には関与していなかった。
 また、p53機能不良群はp53機能良好群に比較して、解剖学的亜部位にかかわらず、外科的に十分な断端距離で切除されていても局所無再発生存率が低かった。

【結論】
 以上より、FFPE検体からのTP53変異解析とp53蛋白表現型を併用したp53機能分類は、p16陽性中咽頭癌を除く頭頸部扁平上皮癌において独立した予後因子であり、局所浸潤性を予測することで、精度の高い予後解析を可能にすると考えられた。今後さらなる技術革新により、術前の微細な生検検体からゲノム解析が可能となり、腫瘍の分子生物学的な特徴を予測することが可能になると考えられる。これらの情報は術式決定の一要素になり、個別化外科治療の樹立につながる可能性がある。

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