食道癌におけるアダプター蛋白ＸＢ１３０とシスチン輸送体ｘＣＴの分子間相互作用解析

2012 Fiscal Year Research-status Report

• Project/Area Number: 24791440
• Research Institution: Kyoto Prefectural University of Medicine
• Principal Investigator: 塩崎 敦 京都府立医科大学, 医学(系)研究科(研究院), 助教 (40568086)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Keywords: アダプター蛋白 / トランスポーター / 食道癌 / 細胞周期

Research Abstract

まず、各種ヒト食道癌細胞株におけるXB130、xCTの蛋白発現レベルを確認した。ウェスタンブロット法により、TE2,TR5,TE9,TE13,KYSE170において両者の発現を確認した。TE13,KYSE170において、xCT-siRNAの導入によりXB130の蛋白・mRNA発現レベルが抑制された。また、TE2,TE9,TE13,KYSE170において、XB130-siRNAの導入によりxCTのmRNA発現レベルが増強され、両者の分子間相互作用の存在が強く示唆された。さらに、TE9,KYSE170にxCTの発現ベクターを導入し、その発現増強効果を確認するとともに、細胞周期・細胞増殖への影響を解析した。現在、両者の直接的結合の有無を確認するため、xCT発現ベクターを導入したTE9,KYSE170細胞株において、Halo tag Resinを用いた共免疫沈降を施行中である。また、shRNA導入により恒常xCT非発現TE13,KYSE170細胞株を樹立中である。一方、食道癌細胞株におけるxCTの細胞増殖制御機構解析を進め、xCT-siRNA導入TE13細胞株のマイクロアレイ解析により、G1-S期関連遺伝子の発現が制御されることを解明した。さらに、食道癌切除検体の免疫組織染色により、xCT発現とKi67・CyclinD1・p21発現の相関解析を行うとともに、xCT高発現が独立した予後因子となることを明らかにした。これらの研究成果は英文雑誌に投稿済みである。同時に、食道癌におけるXB130の細胞周期制御機構と臨床病理学的意義の解明(Ann Surg Oncol. 2012)を行うとともに、他の癌細胞株を用いたXB130に関する基礎実験を行い、Aktを介するシグナル制御機構(PLoS One. 2012)、microRNA発現制御機構(PLoS One. 2013)を新たに解明した。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

当初の研究計画のうち、ヒト食道癌細胞株におけるXB130・xCT発現レベルの解析、一方の発現制御系を用いた際の他者の蛋白・mRNA発現解析による相互作用の有無の検討は、ほぼ終了している。また、各々の因子の食道癌・他癌細胞における細胞増殖・シグナル経路・microRNA制御機構の解析も進展しており、研究成果は既に国内外の学会で発表し、英文雑誌にも投稿・掲載されている。xCT発現ベクター導入食道癌細胞株を用いた共免疫沈降、恒常xCT非発現食道癌細胞株の樹立も進行しており、研究目的・研究計画はおおむね順調に進展していると考える。

Strategy for Future Research Activity

今後、恒常xCT非発現食道癌細胞株、xCT発現ベクターを用いたXB130・xCTの分子間相互作用の解析、細胞周期制御機構の解明をさらに進めていく。また、ヒト食道癌組織におけるXB130、xCT発現レベルの相関性と、臨床病理学的因子との関係の解析を行う。さらに、マウス皮下腫瘍モデルを用いたin vivoでのXB130/xCT分子間相互作用の解析を行っていく予定である。

Expenditure Plans for the Next FY Research Funding

次年度は、細胞培養関連（培地およびウシ胎児血清等）、分子生物学的、生化学的手法に用いる消耗品と、実験動物費用を中心に研究費を使用していく予定である。

Research Products

• [Journal Article] XB130, a new adaptor protein, regulates expression of tumor suppressive microRNAs in cancer cells. (2013)
  • Author(s): Takeshita H, Shiozaki A, Bai XH, Iitaka D, Kim H, Yang BB, Keshavjee S, Liu M.
  • Journal Title: PLoS One Volume: 8 Pages: e59057
  • DOI: 10.1371/journal.pone.0059057.
  • Peer Reviewed
• [Journal Article] Anti-Phosphohistone H3 as an independent prognostic factor in human esophageal squamous cell carcinoma. (2013)
  • Author(s): Nakashima S, Shiozaki A, Ichikawa D, Komatsu S, Konishi H, Iitaka D, Kubota T, Fujiwara H, Okamoto K, Kishimoto M, Otsuji E.
  • Journal Title: Anticancer Res Volume: 33 Pages: 461-467
  • Peer Reviewed
• [Journal Article] A case involving long-term survival after esophageal cancer with liver and lung metastases treated by multidisciplinary therapy: report of a case. (2013)
  • Author(s): Iitaka D, Shiozaki A, Fujiwara H, Ichikawa D, Okamoto K, Komatsu S, Murayama Y, Ikoma H, Kuriu Y, Nakanishi M, Ochiai T, Kokuba Y, Sonoyama T, Otsuji E.
  • Journal Title: Surg Today Volume: 43 Pages: 556-561
  • DOI: 10.1007/s00595-012-0240-7.
  • Peer Reviewed
• [Journal Article] The expression and role of Aquaporin 5 in esophageal squamous cell carcinoma. (2013)
  • Author(s): Shimizu H, Shiozaki A, Ichikawa D, Fujiwara H, Konishi H, Ishii H, Komatsu S, Kubota T, Okamoto K, Kishimoto M, Otsuji E.
  • Journal Title: J Gastroenterol Volume: 印刷中 Pages: 印刷中
  • DOI: 10.1007/s00535-013-0827-9
  • Peer Reviewed
• [Journal Article] Novel technique for dissection of subcarinal and main bronchus lymph nodes using a laparoscopic transhiatal approach for esophageal cancer (2013)
  • Author(s): Shiozaki A, Fujiwara H, Konishi H, Morimura R, Murayama Y, Komatsu S, Kuriu Y, Ikoma H, Kubota T, Nakanishi M, Ichikawa D, Okamoto K, Sakakura C, and Otsuji E
  • Journal Title: Anticancer Res Volume: 印刷中 Pages: 印刷中
  • Peer Reviewed
• [Journal Article] XB130 as an Independent Prognostic Factor in Human Esophageal Squamous Cell Carcinoma. (2012)
  • Author(s): Shiozaki A, Kosuga T, Ichikawa D, Komatsu S, Fujiwara H, Okamoto K, Iitaka D, Nakashima S, Shimizu H, Ishimoto T, Kitagawa M, Nakou Y, Kishimoto M, Liu M, Otsuji E.
  • Journal Title: Ann Surg Oncol Volume: 印刷中 Pages: 印刷中
  • DOI: 10.1245/s10434-012-2474-4
  • Peer Reviewed
• [Journal Article] XB130 mediates cancer cell proliferation and survival through multiple signaling events downstream of Akt (2012)
  • Author(s): Shiozaki A, Shen-Tu G, Bai X, Iitaka D, De Falco V, Santoro M, Keshavjee S, Liu M.
  • Journal Title: PLoS One Volume: 7 Pages: e43646
  • DOI: 10.1371/journal.pone.0043646.
  • Peer Reviewed
• [Journal Article] Claudin 1 mediates TNFα-induced gene expression and cell migration in human lung carcinoma cells. (2012)
  • Author(s): Shiozaki A, Bai XH, Shen-Tu G, Moodley S, Takeshita H, Fung SY, Wang Y, Keshavjee S, Liu M.
  • Journal Title: PLoS One Volume: 7 Pages: e38049
  • DOI: 10.1371/journal.pone.0038049.
  • Peer Reviewed
• [Journal Article] Perioperative outcomes of esophagectomy preceded by the laparoscopic transhiatal approach for esophageal cancer. (2012)
  • Author(s): Shiozaki A, Fujiwara H, Murayama Y, Komatsu S, Kuriu Y, Ikoma H, Nakanishi M, Ichikawa D, Okamoto K, Ochiai T, Kokuba Y, Otsuji E.
  • Journal Title: Dis Esophagus Volume: 印刷中 Pages: 印刷中
  • DOI: 10.1111/j.1442-2050.2012.01439.x.
  • Peer Reviewed
• [Journal Article] Posterior mediastinal lymph node dissection using the pneumomediastinum method for esophageal cancer. (2012)
  • Author(s): Shiozaki A, Fujiwara H, Murayama Y, Komatsu S, Kuriu Y, Ikoma H, Nakanishi M, Ichikawa D, Okamoto K, Ochiai T, Kokuba Y, Otsuji E.
  • Journal Title: Esophagus Volume: 9 Pages: 58-64
  • DOI: 10.1007/s10388-011-0304-6
  • Peer Reviewed
• [Journal Article] Hand-assisted laparoscopic transhiatal approach for mediastinal esophageal duplication cyst resection. (2012)
  • Author(s): Shiozaki A, Fujiwara H, Murayama Y, Komatsu S, Kuriu Y, Ikoma H, Nakanishi M, Ichikawa D, Okamoto K, Ochiai T, Otsuji E.
  • Journal Title: Esophagus Volume: 9 Pages: 247-251
  • DOI: 10.1007/s10388-012-0335-7
  • Peer Reviewed
• [Journal Article] Video-assisted surgery for gastric carcinoma arising in a gastric tube reconstructed retrosternally. (2012)
  • Author(s): Shiozaki A, Fujiwara H, Ichikawa D, Okamoto K, Komatsu S, Murayama Y, Ikoma H, Kuriu Y, Nakanishi M, Ochiai T, Kokuba Y, Sonoyama T, Otsuji E.
  • Journal Title: Surg Today Volume: 42 Pages: 209-213
  • DOI: 10.1007/s00595-011-0029-0.
  • Peer Reviewed
• [Journal Article] Risk factors for postoperative respiratory complications following esophageal cancer resection. (2012)
  • Author(s): Shiozaki A, Fujiwara H, Okamura H, Murayama Y, Komatsu S, Kuriu Y, Ikoma H, Nakanishi M, Ichikawa D, Okamoto K, Ochiai T, Kokuba Y, Otsuji E.
  • Journal Title: Oncol Lett Volume: 3 Pages: 247-251
  • DOI: 10.3892/ol.2012.589
  • Peer Reviewed
• [Journal Article] Blockade of chloride ion transport enhances the cytocidal effect of hypotonic solution in gastric cancer cells. (2012)
  • Author(s): Iitaka D, Shiozaki A, Ichikawa D, Kosuga T, Komatsu S, Okamoto K, Fujiwara H, Ishii H, Nakahari T, Marunaka Y, Otsuji E.
  • Journal Title: J Surg Res Volume: 176 Pages: 524-234
  • DOI: 10.1016/j.jss.2011.10.039.
  • Peer Reviewed
• [Journal Article] Enhancement of the cytocidal effects of hypotonic solution using a chloride channel blocker in pancreatic cancer cells. (2012)
  • Author(s): Nako Y, Shiozaki A, Ichikawa D, Komatsu S, Konishi H, Iitaka D, Ishii H, Ikoma H, Kubota T, Fujiwara H, Okamoto K, Ochiai T, Nakahari T, Marunaka Y, Otsuji E.
  • Journal Title: Pancreatology Volume: 12 Pages: 440-448
  • DOI: 10.1016/j.pan.2012.08.003.
  • Peer Reviewed
• [Journal Article] Ki-67 labeling index as an independent prognostic factor in human esophageal squamous cell carcinoma. (2012)
  • Author(s): Sasagawa H, Shiozaki A, Iitaka D, Ichikawa D, Komatsu S, Fujiwara H, Okamoto K, Nakashima S, Kinoshita O, Murayama Y, Kuriu Y, Ikoma H, Nakanishi M, Ochiai T, Kokuba Y, Otsuji E.
  • Journal Title: Esophagus Volume: 9 Pages: 195-202
  • DOI: 10.1007/s10388-012-0336-6
  • Peer Reviewed
• [Journal Article] Predictive factors for early recurrence in patients with esophageal squamous cell carcinoma after curative esophagectomy. (2012)
  • Author(s): Shimizu H, Shiozaki A, Fujiwara H, Komatsu S, Ichikawa D, Okamoto K, Murayama Y, Kuriu Y, Ikoma H, Nakanishi M, Ochiai T, Kokuba Y, Otsuji E.
  • Journal Title: Esophagus Volume: 9 Pages: 17-24
  • DOI: 10.1007/s10388-011-0308-2
  • Peer Reviewed
• [Journal Article] Intrathoracic supercharge technique for esophageal reconstruction using colon interposition via a retrosternal route. (2012)
  • Author(s): Hino H, Shiozaki A, Fujiwara H, Komatsu S, Ichikawa D, Okamoto K, Murayama Y, Kuriu Y, Ikoma H, Nakanishi M, Ochiai T, Kokuba Y, Otsuji E.
  • Journal Title: Esophagus Volume: 9 Pages: 234-238
  • DOI: 10.1007/s10388-012-0323-y
  • Peer Reviewed
• [Journal Article] Spontaneous rupture of the esophagus with extensive bowel necrosis caused by nonocclusive mesenteric ischemia: report of a case. (2012)
  • Author(s): Nakashima S, Shiozaki A, Fujiwara H, Murayama Y, Komatsu S, Ikoma H, Kuriu Y, Nakanishi M, Ichikawa D, Okamoto K, Ochiai T, Kokuba Y, Otsuji E.
  • Journal Title: Surg Today Volume: 42 Pages: 1107-1110
  • DOI: 10.1007/s00595-012-0195-8.
  • Peer Reviewed
• [Journal Article] A case of a superficial carcinoma of the esophagus with isolated lymph node metastasis around the abdominal aorta. (2012)
  • Author(s): Hatakeyama T, Shiozaki A, Fujiwara H, Ichikawa D, Okamoto K, Komatsu S, Murayama Y, Ikoma H, Kuriu Y, Nakanishi M, Ochiai T, Kokuba Y, Sonoyama T, Otsuji E.
  • Journal Title: Surg Today Volume: 42 Pages: 676-680
  • DOI: 10.1007/s00595-012-0148-2.
  • Peer Reviewed
• [Presentation] Roles of chloride ion transporter in hypotonic shock on gastric cancer cells (2012)
  • Author(s): Atsushi Shiozaki, Daisuke Iitaka, Daisuke Ichikawa, Shuhei Komatsu, Kazuma Okamoto, Hitoshi Fujiwara, Takashi Nakahari, Yoshinori Marunaka, Hisakazu Yamagishi, Eigo Otsuji
  • Organizer: 38th Biennial World Congress of the International College of Surgeons
  • Place of Presentation: Brisbane
  • Year and Date: 20121107-20121110
• [Presentation] Enhancement of cytocidal effect of distilled water using chloride channel blocker in pancreatic cancer cells (2012)
  • Author(s): Yoshito Nakou, Atsushi Shiozaki, Daisuke Ichikawa, Shuhei Komatsu, Hisashi Ikoma, Toshiya Ochiai, Takashi Nakahari, Yoshinori Marunaka, Hisakazu Yamagishi, Eigo Otuji
  • Organizer: 38th Biennial World Congress of the International College of Surgeons
  • Place of Presentation: Brisbane
  • Year and Date: 20121107-20121110
• [Presentation] Roles of XB130, a novel adaptor protein, in esophageal squamous cell carcinoma (2012)
  • Author(s): A. Shiozaki, T. Kosuga, H. Fujiwara, D. Ichikawa, S. Komatsu, K. Okamoto, Y. Murayama, Y. Kuriu, H. Ikoma, M. Nakanishi, T. Ochiai, Y. Kokuba, M. Liu, E. Otsuji
  • Organizer: 13th World Congress of the International Society for Diseases of the Esophagus
  • Place of Presentation: Venice
  • Year and Date: 20121015-20121017
• [Presentation] Roles of xCT, cysteine/glutamate transporter, in esophageal squamous cell carcinoma (2012)
  • Author(s): A. Shiozaki, D. Iitaka, H. Fujiwara, D. Ichikawa, S. Komatsu, K. Okamoto, Y. Murayama, Y. Kuriu, H. Ikoma, M. Nakanishi, T. Ochiai, Y. Kokuba, E. Otsuji
  • Organizer: 13th World Congress of the International Society for Diseases of the Esophagus
  • Place of Presentation: Venice
  • Year and Date: 20121015-20121017
• [Presentation] Clinical significance of Ki-67 labeling index in patients with esophageal squamous cell carcinoma (2012)
  • Author(s): A. Shiozaki, D. Iitaka, H. Fujiwara, D. Ichikawa, S. Komatsu, K. Okamoto, Y. Murayama, Y. Kuriu, H. Ikoma, M. Nakanishi, T. Ochiai, Y. Kokuba, E. Otsuji
  • Organizer: 13th World Congress of the International Society for Diseases of the Esophagus
  • Place of Presentation: Venice
  • Year and Date: 20121015-20121017
• [Presentation] Roles of XB130, a novel regulator of PI3K /Akt pathway, in cancer cell proliferation and survival (2012)
  • Author(s): Shiozaki A、Ichikawa D、Komatsu S、Murayama Y、Kuriu Y、Ikoma H、Kubota T、Nakanishi M、Fujiwara H、Okamoto K、Ochiai T、Liu M、Otsuji E
  • Organizer: 日本癌学会学術総会
  • Place of Presentation: 札幌
  • Year and Date: 20120919-20120921
• [Presentation] The role of Claudin-1 as a mediator of TNF-alpha associated with cell migration (2012)
  • Author(s): Hiroki Shimizu, Atsushi Shiozaki, Daisuke Ichikawa, Hirotaka Konishi, Shuhei Komatsu, Takeshi Kubota, Hitoshi Fujiwara, Kazuma Okamoto, Toshiya Ochiai, Kazuyo Naito, Mingyao Liu, Eigo Otsuji
  • Organizer: 日本癌学会学術総会
  • Place of Presentation: 札幌
  • Year and Date: 20120919-20120921
• [Presentation] XB130 regulates expressions of tumor suppressor microRNAs in thyroid cancer cells. (2012)
  • Author(s): Takeshita H, Shiozaki A, Ichikawa D, Komatsu S, Konishi H, Murayama Y, Kuriu Y, Ikoma H, Kubota T, Nakanishi M, Okamoto K, Fujiwara H, Ochiai T, Kokuba Y, Liu M, Otsuji E
  • Organizer: 日本癌学会学術総会
  • Place of Presentation: 札幌
  • Year and Date: 20120919-20120921
• [Presentation] Expression and prognostic association of XB130/p21 in esophageal squamous cell carcinoma patients (2012)
  • Author(s): Nakashima S、Shiozaki A、Ichikawa D、Komatsu S、Konishi H、Kosuga T、Iitaka D、Kubota T、Fujiwara H、Okamoto K、Ochiai T、Liu M、Otsuji E
  • Organizer: 日本癌学会学術総会
  • Place of Presentation: 札幌
  • Year and Date: 20120919-20120921
• [Presentation] Enhancement of cytocidal effect of hypotonic solution using chloride channel blocker in pancreatic cancer cells (2012)
  • Author(s): Nakou Y、Shiozaki A、Ichikawa D、Komatsu S、Konishi H、Ishii H、Ikoma H、Kubota T、Fujiwara H、Okamoto K、Ochiai T、Marunaka Y、Otsuji E
  • Organizer: 日本癌学会学術総会
  • Place of Presentation: 札幌
  • Year and Date: 20120919-20120921
• [Presentation] 腫瘍進展におけるClaudin-1の意義；TNF-αのメディエーターとしての可能性 (2012)
  • Author(s): 清水浩紀、塩崎敦、市川大輔、小松周平、村山康利、栗生宜明、生駒久視、中西正芳、藤原斉、岡本和真、落合登志哉、國場幸均、内藤和世、Mingyao Liu、大辻英吾
  • Organizer: 日本外科学会定期学術集会
  • Place of Presentation: 千葉
  • Year and Date: 20120412-20120414
• [Presentation] 食道扁平上皮癌手術症例における新規アダプター蛋白XB130の発現解析 (2012)
  • Author(s): 中島慎吾、塩崎敦、市川大輔、小松周平、小菅敏幸、村山康利、栗生宜明、生駒久視、中西正芳、岡本和真、藤原斉、落合登志哉、國場幸均、大辻英吾
  • Organizer: 日本外科学会定期学術集会
  • Place of Presentation: 千葉
  • Year and Date: 20120412-20120414
• [Presentation] 食道扁平上皮癌（ESCC）腫瘍増殖に関わるxCTの役割と意義について (2012)
  • Author(s): 飯高大介 塩崎敦 市川大輔 藤原斉 小松周平 岡本和真 村山康利 栗生宜明 生駒久視 中西正芳 落合登志哉 國場幸均 大辻英吾
  • Organizer: 日本外科学会定期学術集会
  • Place of Presentation: 千葉
  • Year and Date: 20120412-20120414
• [Presentation] 胃癌細胞における細胞内Cl-を介した遺伝子発現制御機構の解明 (2012)
  • Author(s): 北川真希、塩崎敦、市川大輔、小松周平、村山康利、栗生宜明、生駒久視、中西正芳、岡本和真、藤原斉、落合登志哉、國場幸均、丸中良典、大辻英吾
  • Organizer: 日本外科学会定期学術集会
  • Place of Presentation: 千葉
  • Year and Date: 20120412-20120414
• [Presentation] 消化器癌におけるクロライド輸送体制御を利用した蒸留水殺細胞効果の増強 (2012)
  • Author(s): 名幸義仁 塩崎敦 市川大輔 小松周平 小菅敏幸 飯高大介 石井博道 村山康利 栗生宜明 生駒久視 中西正芳 岡本和真 藤原斉 落合登志哉 國場幸均 丸中良典 大辻英吾
  • Organizer: 日本外科学会定期学術集会
  • Place of Presentation: 千葉
  • Year and Date: 20120412-20120414