肺癌における免疫染色を用いた新しいＥＧＦＲ遺伝子変異検出法の確立

2013 Fiscal Year Annual Research Report

• Project/Area Number: 23790425
• Research Institution: Kurume University
• Principal Investigator: 河原 明彦 久留米大学, 大学病院, 臨床検査技師 (00469347)
• Keywords: 病理学 / 癌 / 遺伝子検査 / 免疫染色

Research Abstract

我々は「肺癌における免疫染色を用いた新しいEGFR（上皮増殖因子受容体）遺伝子変異検出法の確立」に関して133肺癌患者のデータ解析を行った．免疫染色とDNA遺伝子変異解析の一致率は陽性判定患者94.6%および陰性判定患者90.6%であり，高い一致率を示した．さらにEGFR遺伝子変異抗体を用いて原発巣と播種巣の解析した結果，再発肺癌患者において，（１）耐性遺伝子を伴う肺癌患者，（２）EGFR遺伝子変異のみを伴う肺癌患者，（３）EGFR遺伝子変異を伴わない肺癌患者のように3群に分けることができた．
本検討の中で細胞材料の判定不可能が多いことが判明したため，固定法によるEGFR遺伝子変異およびEGFRタンパク発現の影響を調査した．15%中性緩衝ホルマリンとCytoRich Redに固定したヒト肺癌培養細胞は，EGFR遺伝子変異特異的抗体に強陽性を示したが，CytoRich Blue と95%エタノール固定液は弱陽性であった．SurePath細胞保存液のDNA保持は15%中性緩衝ホルマリンより優れていたが，EGFR遺伝子変異およびFISH解析は，すべての固定液において良好な結果を得た．胸水中の腺癌細胞において，細胞質抗原は固定後40日間経過しても比較的安定していたが，核内抗原はすべての固定液で発現率の減少を示した．CytoRich RedはEGFR遺伝子変異特異的抗体を用いた免疫細胞化学だけでなく，細胞材料を用いたDNA解析にも適していた．
EGFR-TKI治療におけるEGFR遺伝子変異の検出はDNAによる解析が主流であるが，EGFR遺伝子変異抗体を併用したEGFR遺伝子変異検出アルゴリズムは，EGFR-TKI治療を迅速に投与でき，医療費の削減や患者治療に役立つ新しい迅速な検出方法である．そのためには，がん細胞の固定法に充分留意すべきである．

Research Products

• [Journal Article] FOXP3 expression in tumor cells and tumor-infiltrating lymphocytes is associated with breast cancer prognosis. (2013)
  • Author(s): Takenaka M, Seki N, Toh U, Hattori S, Kawahara A, Yamaguchi T, Koura K, Takahashi R, Otsuka H, Takahashi H, Iwakuma N, Nakagawa S, Fujii T, Sasada T, Yamaguchi R, Yano H, Shirouzu K, Kage M.
  • Journal Title: Molecular and Clinical Oncology Volume: 1 Pages: 625-532
  • DOI: 10.3892/mco.2013.107
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• [Journal Article] Y-box binding protein-1 (YB-1) contributes to both HER2/ErbB2 expression and lapatinib sensitivity in human gastric cancer cells. (2013)
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  • Author(s): Kaida H, Toh U, Hayakawa M, Hattori S, Fujii T, Kurata S, Kawahara A, Hirose Y, Kage M, Ishibashi M.
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  • DOI: 10.1097/MNM.0b013e328360d945.
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• [Journal Article] Expression of Monocarboxylate Transporter (MCT)-4 in Colorectal Cancer and its Role: MCT4 Contributes to the Growth of Colorectal Cancer with Vascular Endothelial Growth Factor. (2013)
  • Author(s): Gotanda Y, Akagi Y, Kawahara A, Kinugasa T, Yoshida T, Ryu Y, Shiratsuchi I, Kage M, Shirouzu K.
  • Journal Title: Anticancer research Volume: 33 Pages: 2941-2947
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• [Journal Article] N-myc downstream-regulated gene 1 promotes tumor inflammatory angiogenesis through JNK activation and autocrine loop of interleukin-1α by human gastric cancer cells. (2013)
  • Author(s): Murakami Y, Watari K, Shibata T, Uba M, Ureshino H, Kawahara A, Abe H, Izumi H, Mukaida N, Kuwano M, Ono M.
  • Journal Title: Journal of biological chemistry Volume: 288 Pages: 25025-25037
  • DOI: 10.1074/jbc.M113.472068.
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• [Journal Article] Prognostic Value of EGFR Mutation and ERCC1 in Patients with Non-Small Cell Lung Cancer Undergoing Platinum-Based Chemotherapy. (2013)
  • Author(s): Yamashita F, Azuma K, Yoshida T, Yamada K, Kawahara A, Hattori S, Takeoka H, Zaizen Y, Kawayama T, Kage M, Hoshino T.
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  • DOI: 10.1371/journal.pone.0071356.
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• [Journal Article] A case of metastasizing pleomorphic adenoma in the maxillary bone appearing twenty years after initial resection of pleomorphic adenoma of the hard palate. (2013)
  • Author(s): Akiba J, Harada H, Kawahara A, Todoroki K, Nagata S, Yano H.
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  • DOI: 10.1111/pin.12087.
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• [Journal Article] Erlotinib resistance in lung cancer cells mediated by integrinβ1/Src/Akt-driven bypass signaling. (2013)
  • Author(s): Kanda R, Kawahara A, Watari K, Murakami Y, Sonoda K, Maeda M, Fujita H, Kage M, Uramoto H, Costa C, Kuwano M, Ono M.
  • Journal Title: Cancer research Volume: 15 Pages: 6243-6253
  • DOI: 10.1158/0008-5472.CAN-12-4502.
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• [Journal Article] Phase II study of personalized peptide vaccination for refractory bone and soft tissue sarcoma patients. (2013)
  • Author(s): Takahashi R, Ishibashi Y, Hiraoka K, Matsueda S, Kawano K, Kawahara A, Kage M, Ohshima K, Yamanaka R, Shichijo S, Shirouzu K, Itoh K, Sasada T.
  • Journal Title: Cancer science Volume: 104 Pages: 1285-1294
  • DOI: 10.1111/cas.12226.
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• [Journal Article] Treatment outcome in patients with stage III breast cancer treated with neoadjuvant chemotherapy. (2013)
  • Author(s): Takahashi R, Toh U, Iwakuma N, Mishima M, Fujii T, Takenaka M, Koura K, Seki N, Kawahara A, Kage M, Ogo E, Shirouzu K.
  • Journal Title: Experimental and therapeutic medicine Volume: 6 Pages: 1089-1095
  • DOI: 10.3892/etm.2013.1289
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• [Journal Article] Potential usefulness of mucin immunohistochemical staining of preoperative pancreatic biopsy or juice cytology specimens in the determination of treatment strategies for intraductal papillary mucinous neoplasm. (2013)
  • Author(s): Hisaka T, Horiuchi H, Uchida S, Ishikawa H, Kawahara R, Kawashima Y, Akashi M, Mikagi K, Ishida Y, Okabe Y, Nakayama M, Naito Y, Yano H, Taira T, Kawahara A, Kage M, Kinoshita H, Shirozu K.
  • Journal Title: Oncology Reports Volume: 30 Pages: 2035-2041
  • DOI: 10.3892/or.2013.2720.
  • Peer Reviewed
• [Presentation] 中間型松果体実質腫瘍の1例． (2013)
  • Author(s): 安部秀幸，河原明彦，杉田保雄，山口知彦，多比良朋希，吉田友子，髙瀨頼妃呼，福満千容，鹿毛政義．
  • Organizer: 第29回日本臨床細胞学会福岡県支部総会
  • Place of Presentation: 北九州
  • Year and Date: 20131201-20131201
• [Presentation] 腫瘍性筋上皮細胞が関与する唾液腺腫瘍の診断アプローチ．第52回 (2013)
  • Author(s): 12. 河原明彦，原田博史，多比良朋希，山口知彦，安部秀幸，吉田友子，髙瀨頼妃呼，福満千容，秋葉 純，鹿毛政義．
  • Organizer: 第52回日本臨床細胞学会秋期大会
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• [Presentation] ワルチン腫瘍と粘表皮癌におけるp63の発現比較． (2013)
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  • Organizer: 第52回日本臨床細胞学会秋期大会
  • Place of Presentation: 大阪
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• [Presentation] Integrin beta1/Src/Akt-driven Bypass Signaling Pathway confers EGFR-TKI Resistance in Lung Cancer Cells. (2013)
  • Author(s): Mayumi Ono, Rina Kanda, Kosuke Watari, Kahori Sonoda, Masashi, Maeda, Yuichi Murakami, Akihiko Kawahara, Masayoshi Kage, Hidetaka Uramoto, Michihiko Kuwano.
  • Organizer: 72th Annual Meeting of the Japanese Cancer Association
  • Place of Presentation: Yokohama
  • Year and Date: 20131003-20131005
• [Presentation] NDRG1 deficiency suppresses tumor angiogenesis through interference of macrophage activation. (2013)
  • Author(s): Kosuke Watari, Hiroshi Nabeshima, Yuichi Fukunaga, TomohiroShibata , Ai Shinoda, Akihiko Kawahara, Yuichi Murakami, Masayoshi Kage, Michihiko Kuwano, Mayumi Ono.
  • Organizer: 72th Annual Meeting of the Japanese Cancer Association
  • Place of Presentation: Yokohama
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• [Presentation] IL-1α-driven AP-1 (Jun /Fos) activation plays a key role in inflammatory angiogenesis by NDRG1 in gastric cancer. (2013)
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  • Organizer: 72th Annual Meeting of the Japanese Cancer Association
  • Place of Presentation: Yokohama
  • Year and Date: 20131003-20131005
• [Presentation] A case of EML4-ALK-positive lung adenocarcinoma with signet-ring cells. (2013)
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  • Organizer: The 12th Korea-Japan Joint Meeting for Diagnostic Cytopathology.
  • Place of Presentation: Korea
  • Year and Date: 20130907-20130907
• [Presentation] Claudin-1 is detection marker of squamoid corpuscles in pancreatoblastoma. (2013)
  • Author(s): Tomoki Taira, Akihiko Kawahara, Tomohiko Yamaguchi, Hideyuki Abe, Tomoko Yoshida, Yorihiko Takase, Chihiro Fukumitsu, Jun Akiba, Kage Masayoshi.
  • Organizer: The 12th Korea-Japan Joint Meeting for Diagnostic Cytopathology.
  • Place of Presentation: Korea
  • Year and Date: 20130907-20130907
• [Presentation] Integrinβ1/Akt経路活性化を介した肺癌における新規エルロチニブ耐性獲得メカニズム． (2013)
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  • Place of Presentation: 京都
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• [Presentation] Y-box結合タンパク(YB-1)はヒト胃癌においてHER2/ErbB2発現とラパチニブの感受性を制御する (2013)
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  • Organizer: 第17回日本がん分子標的治療学会
  • Place of Presentation: 京都
  • Year and Date: 20130612-20130614
• [Presentation] 唾液腺領域における良性腫瘍を悪性と診断しない鑑別ポイント． (2013)
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  • Organizer: 第54回日本臨床細胞学会総会
  • Place of Presentation: 東京
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• [Presentation] 液状気管支細胞診を用いた肺癌診断． (2013)
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  • Organizer: 第54回日本臨床細胞学会総会
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  • Year and Date: 20130531-20130602
• [Presentation] 細胞診による子宮内膜癌のtype分類の検討． (2013)
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  • Organizer: 第54回日本臨床細胞学会総会
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  • Year and Date: 20130531-20130602
• [Presentation] Glucose transporter-1(GLUT-1) expression in endometrial lesions. (2013)
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  • Organizer: 18th International Congress of Cytology
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